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Expanding global distribution of rotavirus serotype G9: detection in Libya, Kenya, and Cuba.

Serotype G9 may be the fifth most common human rotavirus serotype, after serotypes G1 to G4. In three cross-sectional studies of childhood diarrhea, we have detected serotype G9 rotaviruses for the first time in Libya, Kenya, and Cuba. Serotype G9 constituted 27% of all rotaviruses identified, emphasizing the reemergence of serotype G9 and suggesting that future human rotavirus vaccines will need to protect against disease caused by this serotype.

Child↗

Salmonella enterica serotypes isolated from imported frozen chicken meat in the Canary islands.

To determine the prevalence of Salmonella enterica serotypes in imported frozen chicken meat, 406 samples (whole chicken, legs, and breast meat) were analyzed for Salmonella according to ISO6579 rules, serotypes were assigned, and phage typing was conducted for Salmonella serotypes Enteritidis, Typhimurium, and Heidelberg. The overall frequency of Salmonella isolation was 16.5%. By country of origin, the highest percentage of cases was found among the samples from France followed by samples from Brazil. The differences between legs and breast meat were significant. The most frequently isolated serotype of Salmonella was Enteritidis, followed by Salmonella Heidelberg, Salmonella Typhimurium, and Salmonella Virchow. By country of origin, we identified a large percentage of serotype Salmonella Enteritidis in the samples imported from Brazil. There was a greater diversity of serotypes isolated from the French samples, and Salmonella Enteritidis was not the dominant strain. In the samples from the United States, the only serotype isolated was Salmonella Kentucky, although a smaller number of samples was analyzed. The Salmonella Enteritidis phage type that prevailed in both France and Brazil was 4. Phage types 204c and 204 were identified for Salmonella Typhimurium, and phage types 8, 31, and 37 were identified for Salmonella Virchow.

Animals↗

Growth and stress resistance variation in culture broth among Listeria monocytogenes strains of various serotypes and origins.

Twenty-five Listeria monocytogenes strains of various serotypes and sources, including clinical and food isolates associated with the same outbreaks, were characterized and compared based on growth rates and heat and acid death rates. Growth was monitored in tryptic soy broth supplemented with 0.6% yeast extract (TSBYE) at 4 and 30 degrees C for 32 days and 20 h, respectively. Heat and acid stress responses in TSBYE heated to 55 degrees C or acidified to pH 3.0 with lactic acid were evaluated for 240 or 120 min, respectively. Extensive variation in growth and stress resistance was observed among the tested strains. Growth rate differences were less evident at 30 than at 4 degrees C, where growth rates (log CFU per milliliter per day) ranged from 0.28 to 0.43. Thermal and acid death rates (log CFU per milliliter per minute) ranged from -0.023 to -0.052 and from -0.012 to -0.134, respectively. Serotype appeared to play a significant role (P < 0.05) only with respect to the heat resistance of the organism. Serotype 4b isolates as a group had lower heat resistance than did isolates representing all other serotypes combined. Although no clear origin-related (food versus clinical) trends were observed under the tested conditions, outbreak-related isolates of serotype 4b had lower acid death rates (higher acid resistance) (P < 0.05) than did the rest of the strains belonging to this serotype. Strain Scott A exhibited slow growth at 4 degrees C and low acid resistance, behavior that was distinct among both clinical and serotype 4b isolates. The results of this study highlight the risks associated with extrapolation to other strains of findings obtained with only one strain of L. monocytogenes. This information should be useful when test strains are to be selected for the evaluation of antimicrobial alternatives in ready-to-eat meat and other food products and when risk assessments are to be conducted.

Adaptation, Physiological↗

Serotypes of Pasteurella haemolytica and Pasteurella trehalosi isolated from farm animals in Hungary.

The biochemical and serological characteristics of 486 P. haemolytica and 31 P. trehalosi strains (517 in total) isolated from different lesions of cattle, sheep, goats, pigs and poultry were examined. A total of 476 P. haemolytica strains (97.9%) showed the characteristics typical of the former biotype A of P. haemolytica, while 10 isolates (2.1%), all from poultry, could not be biotyped. A total of 481 strains (93.0%) could be assigned to one of the 17 serotypes of P. haemolytica-P. trehalosi and 36 strains (7.0%) could not. The majority (83.6%) of the cattle isolates were serotypes A1 and A2. Among strains isolated from sheep all serotypes of P. haemolytica could be identified with the exception of A14, but serotypes A1, A2, A6, A8 and A5 were the most frequent. The overwhelming majority (94%) of the caprine isolates were A2, other serotypes occurred only sporadically. The pig isolates, which could be isolated only very rarely, represented different serotypes, while none of the 10 strains isolated from poultry could be biotyped or serotyped.

Animals↗

Characterisation of a "European-like" serotype G8 human rotavirus isolated in Australia.

An atypical human rotavirus strain, DG8, was isolated from a 13-month-old child hospitalised with acute gastro-enteritis in Australia. The virus could not be serotyped by enzyme immunoassay (EIA) using standard reagents specific for common Group A human rotavirus G serotypes. The deduced amino acid sequence of the outer capsid glycoprotein, VP7, indicated that this strain belonged to the uncommon human serotype G8. This was confirmed by EIA incorporating a G8-specific neutralising monoclonal antibody (NMAb). The VP4 genotype of DG8 was determined as P[14], equivalent to P serotype P3B, by sequence analysis and confirmed by EIA incorporating a P3B-specific NMAb. Electrophoretic analysis of DG8 genomic dsRNA indicated that the virus exhibited a "long" electropherotype. Northern hybridisation analysis (using a whole-genome probe derived from DG8) indicated that DG8 shared overall homology with the European serotype G8 strain, HAL1166 (11 of 11 genes). In contrast, only 9 of 11 genes of DG8 hybridised with the Asian serotype G8 strain, B37, and with the bovine G8 strain, A5. Hence, DG8 displayed features reminiscent of the human serotype G8 rotaviruses isolated in Europe in the mid-1980s rather than the geographically local G8 Asian strains isolated a decade earlier. It is possible that DG8 arose through reassortment between human and bovine rotaviruses.

Amino Acid Sequence↗

[Monoclonal antibodies to dengue virus 4: the spectrum of the reactivity to 4 serotypes of dengue].

These types of monoclonal antibodies 8E8, 3F7 and 1E9 to dengue 4 virus H-241 strain. These monoclonal antibodies show various patterns of reactivity to the four dengue serotypes and different antigen preparations of serotype 4 when they were tested in various serological methods. The monoclonal antibody 8E8 exhibited a specificity of serotype (D-2; by hemagglutination inhibition); subcomplex (D-2 and D-4 by immunofluorescence) and complex (by immunoperoxidase technique). It was able to neutralize by 80% homologous virus and it turned out to be the only reactive monoclonal antibody in the complement fixation test. The monoclonal 3F7 did not react to by hemagglutination inhibition, recognized serotypes D-1, D-2, D-3 and D-4 by immunofluorescence and only serotypes D1 and D4 by immunoperoxidase technique but it was unable to neutralize the homologous virus. The 1E9 antibody was reactive to serotypes D1, D-2, D-3 and D-4 only by hemagglutination inhibition and neutralized serotype D-4. All the monoclonal antibodies were able to react to various dengue antigens through an ELISA of double antibody and showed fluorescent activity against 38th pass in Beagle dog kidney culture; however, they could not react to a D-4 recombinant antigen.

Aedes↗

Relationship between pathogenesis of Guillain-Barre syndrome and Penner's serotypes of Campylobacter jejuni.

OBJECTIVE: To investigate the relationship between the pathogenesis of Guillain-Barre syndrome (GBS) and Campylobacter jejuni (C. jejuni) in China. METHODS: C. jejuni strains were isolated from fresh stools of 47 GBS patients in Beijing area from 1995 to 1997 by modified Skirrow's method. Serotyping of C. jejuni was performed with Penner's method in 47 GBS patients and 171 patients with C. jejuni enteritis in our hospital during the same period. The stools from which no C. jejuni strains could be isolated were typed by PCR-RFLP (restriction fragment length polymorphism) method. RESULTS: Six C. jejuni strains were isolated by Skirrow's method, including 3 strains of Penner serotype 2, 1 Penner serotype 4, 1 Penner serotype 19 and 1 Penner serotype 26. Three strains of DNA-I, which was equal to the same type of Penner serotype 19, were found by PCR-RFLP method. CONCLUSION: C. jejuni isolated from Beijing area are similar to those reported in literature, which once more confirms the molecular mimicry pathogenetic theory of GBS caused by C. jejuni infection in China. The similarities of serotypes between C. jejuni strains from GBS patients and those from C. jejuni enteritis patients may explain the high incidence of GBS following C. jejuni infection in China.

Campylobacter Infections↗

Differentiation of Shigella strains by plasmid profile analysis, serotyping and phage typing.

Ninety-one Shigella flexneri and 29 Shigella sonnei strains isolated during 1994 from sporadic cases of shigellosis and healthy carriers were analyzed for plasmid profile in order to compare the discriminating ability of this method with that of serotyping and phage typing. Our study revealed 10 plasmid profiles (PP) among S. sonnei strains. A total of 26 out of 29 (89%) S. sonnei isolates could be placed into two phage types (type 1 and 20) comprising four PP for phage type 1 and seven PP for type 20, respectively. Twenty-three different PP were identified among S. flexneri strains. Each serotype was associated with a specific predominant plasmid profile, except serotype 2a. This serotype, the most frequently isolated in Romania, was still rather homogeneous: 33 out of 39 isolates belonged to phage type 125, 27 of which could be placed into two related PP (F10 and F17). Comparison of plasmid patterns of epidemiologically independent S. flexneri serotype 2a isolates with those exhibited by 45 serotype 2a isolates associated to six independent outbreaks revealed the same homogeneity. Thirty-eight strains, representing 4 of 6 outbreaks, had F10 and F17 plasmid patterns. The discrimination indices (D) for plasmid profile analysis alone (D = 0.890) and for the combination of serotyping and phage typing (D = 0.841) indicate that both typing systems have a nearly similar ability of discriminating among S. flexneri strains. By combining the results of the three typing methods, a total of 42 types are distinguished and the D value is 0.942. Our data suggest that plasmid profile analysis can complement phenotyping methods resulting in a degree of discrimination that cannot be achieved by either system alone.

Bacterial Typing Techniques↗

[Use of PCR and nucleotide sequence analysis of the VP7 gene for detection of bovine rotaviruses and identification of their G serotypes].

A Vp7 gene fragment PCR protocol was developed to detect the bovine rotaviruses and to identify their G serotypes. The most widespread bovine rotaviruses of G serotypes (G6, G8 and G10) can be distinguished on the basis of the PCR fragment size, while other G serotypes can be differentiated through a comparative analysis of the VP7 gene fragment nuclcotide sequence. Twenty-four bovine rotavirus field isolates were detected, and their G stereotypes were determined by using the method in question. Fourteen isolates were shown to be of G6 serotype; four of them were of G8, five--of G10, and one isolate was of G11 serotype. A possibility of detecting more than one isolate by this method was shown. Finally, a feasibility of using the method for searching for rotaviruses of new bovine rotavirus G serotypes and for rotaviruses, which do not belong to a so far described G serotypes, is discussed.

Animals↗

Serotype specificity of vaccine-induced immunity to pertussis.

Two large epidemics of pertussis occurred in Britain during 1977-79 and 1981-83, following a decline in vaccine uptake in the mid-1970s. Serotype 1,3 predominated during both epidemics but serotypes 1,2 and 1,2,3 constituted 32% (243 of 769) of the isolates from unvaccinated children compared with only 10% (9 of 86) of the isolates from vaccinated children (p < 0.005). Whole-cell vaccines, although effective against infection with all serotypes, have a higher efficacy against serotypes 1,2 and 1,2,3 than against serotype 1,3. These data suggest that not only should acellular vaccines contain agglutinogens specific for each serotype but trials should demonstrate efficacy against all three serotypes.

Bordetella pertussis↗

[O-serotypes, biotypes, and antimicrobial susceptibility of Serratia marcescens isolates from clinical specimens: 3rd report].

O-serotypes, biotypes, and drug resistance of Serratia marcescens strains isolated from various clinical specimens in Showa University Fujigaoka Hospital were investigated: period I (1979. 1-1982.3), 122 strains; period II (1983. 1-1985. 3), 198; period III (1985. 4-198. 3), 129; period IV (1989. 1-1991. 3), 99. The frequency of serotype O 3 was higher than those of the other serotypes after the period II and ranged 17 to 37%. The isolation frequency (27%) of serotype O 4 was higher than those of other serotypes in the period I. However, this frequency decreased in the period II to III and, in the period IV (10%), was higher than in the period III. The isolation frequency of serotype O 17 suddenly increased in the period II (27%) alone, and 42% of the biotypes, obtained with API 20E, showed 5305701. The isolation frequency of non-typable strains ranged 9 to 14% through all periods. The group including 5317721 and 5307721 clustered by the furthest neighbor method, more frequently appeared through all periods. The former code of urease positive more frequently appeared in the period II to IV. The isolation frequency of the latter code of urease negative reduced to a minimum (18%) in period II, though it increased in the period II to IV. The isolation frequency (22%) of the pigmented strains in the period IV increased much higher than in the period I to III (ranged 2 to 4%). The frequency of the pigmented strains of serotype O 3, not belonged to the Grimont's biotype, was 27% of all pigmented strains isolated in the period IV.(ABSTRACT TRUNCATED AT 250 WORDS)

Drug Resistance, Microbial↗

Use of plasmid profiles to differentiate strains within specific serotypes of classical enteropathogenic Escherichia coli.

1. The usefulness of plasmid profile analysis to differentiate strains of enteropathogenic Escherichia coli (EPEC) was evaluated by studying 123 strains of the most prevalent serotypes causing infant diarrhea in the city of São Paulo, Brazil, i.e., O111ab:H-, O111ab:H2 and O119:H6. 2. No common profiles were found among strains of distinct serotypes. However, within each serotype, most of the strains were grouped within a few major profiles. More than 68% of the strains of serotypes O111ab:H- and O111ab:H2 were included in 6 and 9 major profiles, respectively. In serotype O119:H6, about 48% of the strains were included in 3 major profiles. 3. This analysis suggests that only a few EPEC clones are causing infant diarrhea in São Paulo and revealed that the distribution of serotypes O111ab:H- and O111ab:H2 during the one-year study was at least partly determined by small outbreaks of the most common profiles. 4. We conclude that plasmid profile analysis is very useful to differentiate strains within specific EPEC serotypes.

Brazil↗

[Prevalence of serotypes and antimicrobial resistance of invasive strains of Streptococcus pneumoniae].

OBJECTIVE: To determine the prevalence of serotypes and antimicrobial susceptibility of invasive strains of Streptococcus pneumoniae and to discuss the implications of these findings for vaccine formulation. METHODS: Strains of Streptococcus pneumoniae obtained from normally sterile fluids from patients admitted with invasive diseases were isolated and identified at the Hospital de Clínicas, Universidade Federal de Uberlândia, state of Minas Gerais, and forwarded to Instituto Adolfo Lutz, state of São Paulo, for further identification, serotyping and determination of antimicrobial susceptibility. RESULTS: From April 1999 to March 2003, 148 invasive strains of Streptococcus pneumoniae were obtained. The age of patients ranged from 1 day to 88.83 years (mean: 21.33+/-25.82 years; median: 4.42 years). Eighty-four (56.7%) patients were male. The most common diagnoses were pneumonia (91 cases; 61.4%), meningitis (32 cases; 21.6%) and occult bacteremia (15 cases; 10.1%). Strains were isolated mostly from blood (76 occasions; 51.3%), pleural fluid (39 occasions; 26.3%) and cerebrospinal fluid (30 occasions; 20.2%). There were 23 different serotypes, and the most common were 14, 3, 1, 5, 6A, 6B and 18C. Among 30 (20.2%) oxacillin-resistant strains, 23 (15.5%) were confirmed as resistant to penicillin (12.8% intermediate resistance and 2.7% full resistance). Oxacillin-resistant strains were restricted to serotypes 14, 23F, 19A and 6B. Resistance to penicillin varied with age, being more common in children under two years of age (p = 0.0008). We observed decreased sensibility to sulfamethoxazole-trimethoprim (92 isolates; 63.4%), to erythromycin (12 isolates; 8.3%), to clindamycin (12 isolates; 8.7%), to ofloxacin (one strain; 0.8%) and to cefotaxime (three strains; 2%; also resistant to penicillin). All isolates were susceptible to chloramphenicol, rifampin and vancomycin. CONCLUSIONS: The decreased susceptibility to penicillin, detected in 15.5% of the strains was predominant in children under two years of age. There were 23 different Streptococcus pneumoniae serotypes. The 23-valent polysaccharide vaccine covers 82.6% of the serotypes and 90.2% of the invasive strains isolated in this population. In addition, 46.7% of the serotypes and 63.6% of the strains isolated from children until five years of age are covered in the currently available 7-valent conjugated vaccine (PN CRM7).

Adolescent↗

[Prediction of serotypes of dengue virus by response to IgM antibodies].

A study of 4 groups of serum samples from Cuba, Costa Rica, Nicaragua and Panama was made to determine the serotypes of dengue virus causing epidemics or outbreaks by using IgM antibody-capture ELISA(MAC-ELISA). Dengue-IgM diagnostic kit was independently used in each of the serotypes and an index value was calculated (optical density of the sample/cut-off value) in each case, with which two statistical analyses were made: Fisher's variances and Litter significant difference (LSD) calculation. Such calculations allowed determining that in Cuba and Panama, serotype 2 circulated whereas in Costa Rica, serotype 1 prevailed. These were the isolated serotypes in various epidemics. Regarding Nicaragua, very heterogeneous responses were obtained, the responsible serotype could not be determined since more than two serotypes co-circulated in this country.

Dengue Virus↗

[Analysis of VP7 serotype of rotavirus in infantile diarrhea in Hangzhou area from 2001 to 2003].

OBJECTIVE: To investigate molecular epidemiologic features of rotavirus (RV) infection in infantile diarrhea in Hangzhou area. METHODS: Stool specimens of 683 infants with suspected acute viral enteritis in the autumn and winter of 2001 - 2003 were collected. RV (group A) was detected by using latex agglutination test (LAT). VP7 serotype (G) positive specimens were detected by using enzyme linked immunosorbent assay (ELISA) and then the RNA of the virus was determined with reverse transcription polymerase chain reaction (RT-PCR). cDNA of VP7 gene fragment was sequenced by automatic gene analyzor (ABI3730) and compared with the RV VP7 gene sequences stored in Genebank. RESULTS: RV was detected in 297 of 683 (43.5%) specimens by LAT. The highest frequency of RV (group A) detected was 52.9% (228/431) in patients aged 7 - 18 months. The prevalent serotypes were G1 (36.7%, 109/297) and G3 (30.9%, 92/297), followed by mixed type (11.8%, 35/297), untyped (9.4%, 28/297), G4 (7.1%, 21/297) and G2 (4.0%, 12/297). The prevalent serotypes seen each year were different. G1 (54.9%, 45/82) was the major serotype in 2001 followed by G3 (14.6%, 12/82). In 2003, the major serotype was G3 (43.0%, 63/146) and followed by G1 (29.5%, 43/146). The reliability of ELISA was confirmed by RT-PCR, gene sequencing and homology analysis. CONCLUSION: The main prevalent serotypes of VP7 of rotavirus were G1 and G3. The dominant serotypes of rotavirus varied in Hangzhou area from 2001 to 2003.

Antigens, Viral↗

[Antimicrobial susceptibility and serotype distribution of pneumococci strains isolated from meningitis patients].

Treatment of life threatening pneumococcal infections such as meningitis has recently become problematic due to the emergence of antibiotic resistant strains. Antimicrobial susceptibility data usually derived from the studies that included all clinical pneumococcal isolates. However, resistance patterns of this microorganism isolated from meningitis cases in our country are not exactly known. The aim of this study was to determine the antimicrobial susceptibility and serotypes of Streptococcus pneumoniae strains isolated from meningitis cases. This retrospective study was designed in three university hospitals in Turkey and 72 pneumococci isolated from patients with meningitis were evaluated. In this study disk diffusion test and E-test methods were used to determine the susceptibility of pneumococci to some antibiotics. All S. pneumoniae isolates were serotyped using Quellung reaction. Although resistance for oxacillin was found by disc diffusion method in 11 isolates, only six of them were found to be resistant by E-test. By the latter procedure, no resistance was recorded against ceftriaxone and meropenem, while chloramphenicol resistance was found as 1.4%. In our study, erythromycin, ciprofloxacin and levofloxacin resistance were 2.8%, TMP-SMX resistance was 26.4%, while no vancomycin resistance was detected by disk diffusion. In evaluation of 72 pneumococci, we found 16 different serotypes and four isolates could not be serotyped. The serogroup 23 (n: 19) was the most common one followed by serotype 19 (n: 9) and serotype 14 (n: 7). Of six resistant isolates, three pneumococci were serogroup 23 and the remaining were from three different serotype/serogroups 11, 14 and 19. As a result penicillin resistance in pneumococci isolated from meningitis was low and there was no resistance to ceftriaxone. It seems that ceftriaxone is an appropriate choice for empirical treatment of meningitis in our patients. These findings also revealed that pneumococcal polysaccharide capsule vaccines in use cover most of the invasive pneumococcal serotypes.

Adolescent↗

Isolation of monoclonal antibodies-escape variant of dengue virus serotype 1.

INTRODUCTION: During an outbreak from December 2004 to March 2005, 138 isolates of dengue virus were prospectively obtained from acute-phase serum samples of 1,067 patients with the provisional clinical diagnosis of acute dengue illness admitted to the adult wards of Hospital Tengku Ampuan Rahimah, Klang, Malaysia. Of the 138 dengue virus isolates, 87, 11, 24 and 3 were typed as dengue serotypes 1, 2, 3 and 4, respectively, by a commercial dengue virus typing kit using monoclonal antibodies (Mab). 13 dengue virus isolates could not be assigned to any specific serotype by serotyping Mab and molecular typing using dengue-type specific molecular typing primer pairs. We report the associated clinical features and limited molecular genetics of this Mab-escape dengue virus variant. METHODS: Limited molecular characterisation of the Mab-escape dengue virus variants with respect to a few concurrently isolated dengue serotype 1 virus was performed by reverse transcriptase polymerase chain reaction (RT-PCR), followed by nucleic acid sequencing of the 500-bp dengue virus partial genomic capsid-PreM fragment. RESULTS: The aligned nucleic acid sequence of RT-PCR products showed that these Mab-escape variants were of identical nucleic acid sequence, and shared the highest sequence homology (99 percent) with dengue virus serotype 1 (GeneBank accession No. AB178040) isolated from a Japanese patient in 2004. Though these Mab-escape dengue virus variants were noted to replicate to a 2-log higher titre than the current circulating dengue virus serotype 1, there was no significant difference between these variants and the currently circulating dengue virus serotype 1 with respect to disease severity (dengue fever versus dengue haemorrhagic fever) and clinical features. CONCLUSION: There was no significant difference in the proportion of patients developing dengue haemorrhagic fever following acute infection by Mab-escape dengue virus 1 variant in comparison with infection by the conventional dengue virus 1. Similarly, there was no significant difference in the pattern of clinical presentations following acute infection by the two different strains of virus.

Amino Acid Sequence↗

[Distribution of serotypes A and B of Candida albicans in 502 strains isolated from pathological specimens].

BACKGROUND: Candida albicans is distributed in at least two serological groups (A and B) depending on its antigenic structure. These characteristics have been used to investigate the epidemiology of candidiasis. No information is available in this country about the distribution of these serotypes in a general population of patients with candidiasis of different localizations. The aim of the present study was to correlate the frequency of C. albicans serotypes with the origin of the clinical samples. METHODS: In 502 strains of C. albicans isolated from pathological products, serotype was evaluated by means of a latex agglutination test with polyclonal monovalent antisera. A statistical analysis was carried out to investigate whether there was any relationship between the serotype frequency and the origin of the strains. RESULTS: The serotype A of C. albicans represented 78% of all isolates. However, there was a significantly higher relative frequency of serotype B in vaginal and oropharyngeal samples. No differences were found in isolates from AIDS patients. CONCLUSIONS: The serotyping method should be considered a contribution, even if limited, to the knowledge of the epidemiology of Candida albicans infections.

Acquired Immunodeficiency Syndrome↗