Osmotically stabilized media have been reported to increase the recovery rate of various bacteria from blood. This study was made to determine the effect of high concentrations of sucrose on the cultivation of S. typhi from blood. Sucrose in 15% or 30% concentration in the blood culture media retarded the growth. The mean incubation time for the appearance of growth was significantly longer in the media with sucrose. In those blood specimens which rendered growth of S. typhi in both media with and without sucrose, the incubation times were compared; and it was found that the majority of the specimens showed faster growth in the media without sucrose. Experimental cultures showed that the higher the sucrose concentration the lighter and slower were the growths of S. typhi. These tendencies were also observed in the growth of E. coli, P. aeruginosa, S. aureus, beta-hemolytic Streptococcus, alpha-hemolytic Streptococcus and S. pneumoniae.
Culture Media ; Human ; Salmonella typhi/drug effects ; Salmonella typhi/growth & development* ; Salmonella typhi/isolation & purification ; Sucrose/pharmacology*

Female ; Humans ; Infant ; Meningitis, Bacterial ; complications ; Salmonella Infections ; complications ; Salmonella typhi ; isolation & purification ; Sepsis ; complications

During the 8-month period of May to December, 1978, a total of 3,529 blood cultures were taken from Yonsei Medical Center patients and the effect of blind subculture in the initial detection of Salmonella typhi positive culture was analyzed. The blind subculture at the end of 1-day incubation (1-d BS) detected 35.0% of S. typhi positive specimens. All of the S. typhi positive specimens by 1-d BS were a1so macroscopically positive. However, by doing slide agglutination with the growth on subculture plate S. typhi was identifiable tentatively. This saved a day compared to macroscopic examination alone. Therefore the 1-d BS is concluded to be a valuable procedure for the isolation of this organism from blood. For the isolation of S. typhi 7-day incubation was concluded adequete based on the fact that there was only 1 specimen which became positive after over 1-week incubation.
Bacteriological Techniques* ; Blood/microbiology ; Comparative Study ; Culture Media ; Human ; Salmonella typhi/isolation & purification* ; Time Factors

The authors analysed bone marrow findings of sixteen cases of culture proven typhoid fever to reveal the pathologic changes according to the disease stage. The most frequent finding was chronic granulomatous inflammation (eight cases). Infection (bacteria) associated hemophagocytic syndrome (four cases), reactive marrow (two cases), and non specific findings (two cases) were also encountered. Granulocytic hyperplasia with hemophagocytosis appeared at the early stage and was followed by infection (bacteria) associated hemophagocytosis and granuloma in proliferative stage. In lysis (late) stage, granulomatous inflammation was noted. However, resolution of granulomatous inflammation was not distinct. Some nuclear debris and phagocytosis were remarkable in well-formed granulomas. Thrombocytopenia was the most remarkable peripheral blood finding at the time of biopsy. Anemia, leukopenia, and pancytopenia were also observed in descending order.
Adult ; Bone Marrow/microbiology/*pathology ; Female ; Humans ; Male ; Middle Aged ; Salmonella typhi/isolation & purification ; Thrombocytopenia/pathology ; Typhoid Fever/microbiology/*pathology

OBJECTIVETo develop a rapid and definite diagnostic test of bacterial enteritis caused by pathogenic enterobacteria, the most frequent etiologic agent of infectious enteritis in the world.

METHODSA set of conventional PCR assays were applied to detect and identify salmonella, shigella, and E. coli O157:H7 directly from pure culture and fecal samples. The general primers of pathogenic enterobacteria were located on the uidA gene, which were found not only in E. coli nuclear acid, but also in shigella and salmonella genes. Shigella primer was from ipaH gene whose coded invasive plasmid relative antigen existed both in plasmid and in genome. The primers of salmonella were designed from the 16SrRNA sequence. The primer of E. coli O157:H7 was taken from eaeA gene. Five random primers were selected for RAPD. The detection system included common PCR, semi-nested PCR and RAPD.

RESULTSThis method was more sensitive, specific and efficient and its processing was rapid and simple. For example, the method could be used to specifically detect and identify salmonella, shigella, and E. coli O157:H7, and its sensitivity ranged from 3 to 50 CFU, and its detection time was 4 hours.

CONCLUSIONThis PCR method, therefore, can serve as a routine and practical protocol for detecting and identifying pathogenic microorganisms from clinical samples.

DNA Primers ; DNA, Bacterial ; analysis ; Escherichia coli O157 ; isolation & purification ; Feces ; microbiology ; Humans ; Polymerase Chain Reaction ; Salmonella typhi ; isolation & purification ; Sensitivity and Specificity ; Shigella flexneri ; isolation & purification

Objective: To understand the epidemiological and molecular characteristics of typhoid and paratyphoid in China from 2009 to 2013, and provide evidence for the prevention and control of typhoid and paratyphoid, the development and improvement of surveillance strategies. Methods: Epidemiological analysis was conducted on the incidence data of typhoid and paratyphoid, and related public health emergencies in China during 2009-2013. Pathogen isolation and culture, serologic test were conducted for the typhoid and paratyphoid cases from 13 national surveillance sites. The isolates were subjected to antimicrobial susceptibility testing. Pulsed-field gel electrophoresis (PFGE) was performed for the molecular typing of these isolates. Results: The average incidence of typhoid and paratyphoid in China during this period was 1.03/100 000. The reported case number and incidence decreased with year. The provinces reporting high case numbers were Yunnan, Guizhou, Guangxi, Hunan, Zhejiang, Guangdong and Xinjiang. The incidence of age group 0-4 years was highest. The proportion of farmers and children outside child care settings showed an increasing tendency over time. The annual incidence peak was during July-August. Twenty five outbreaks occurred during 2009-2013. The results of pathogen isolation and culture showed that the positive rate was 3.00% (940/31 322), among the positive isolates, the proportion of Salmonella paratyphi A accounted for higher proportion (68.19%, 641/940) compared with Salmonella typhi (31.60%, 297/940). The drug resistances of Salmonella typhi and Salmonella paratyphi varied, but their resistances to nalidixic acid were highest (50.22% and 85.33%) respectively. A certain amount of Salmonella typhi isolates showed the resistance to the 3rd generation cephalosporins. PFGE analysis showed divergent patterns of Salmonella typhi compared with limited patterns of Salmonella paratyphi A. Conclusion: The epidemic level of typhoid and paratyphoid in China was relatively low, but the outbreak occurred occasionally. It is necessary to enhance the laboratory-based surveillance, particularly the capability of etiological diagnosis, outbreak investigation, response and antibiotic resistance monitoring, and conduct risk factor investigation in provinces with high incidences in recent years.
Child ; Child, Preschool ; China/epidemiology* ; Disease Outbreaks ; Drug Resistance, Bacterial/genetics* ; Electrophoresis, Gel, Pulsed-Field ; Epidemics ; Farmers ; Humans ; Incidence ; Infant ; Molecular Typing ; Paratyphoid Fever/microbiology* ; Population Surveillance ; Salmonella paratyphi A/isolation & purification* ; Salmonella typhi/isolation & purification* ; Typhoid Fever/microbiology*

OBJECTIVETo analyses the genetic diversity and relationship of Salmonella typhi strains isolated from different years and districts in Guizhou province.

METHODSRibotyping with 16s rDNA probe was used to describe the diversity of the 209 strains which were isolated in 26 counties of Guizhou province, from 1959 to 1999. The antibiotics resistance was also studied.

RESULTSTwenty-six ribotypes were found in all 209 strains, with two dominant types. The strains isolated from local typhoid epidemics belonged to the unique Ribotypes. The major ribotypes of the resistant strains were RT7 and RT1.

CONCLUSIONThe Salmonella typhi isolates from Guizhou diverged obviously. The abundant clones and multi-resistance of the strains might serve the major reasons of the high morbidity of typhoid in Guizhou.

Blotting, Southern ; China ; DNA, Bacterial ; genetics ; DNA, Ribosomal ; genetics ; Phylogeny ; RNA, Ribosomal, 16S ; genetics ; Ribotyping ; Salmonella typhi ; classification ; genetics ; isolation & purification

This experiment was conducted to assess the efficacy of typhoid vaccine newly produced by purifying Vi antigen of Salmonella typhi. With Karber method, LD50 of challenging organism (S. typhi ty2) was determined as 6.31 CFU/mouse, and then the organism was used for the study. With Probits method, ED50 of the vaccine was determined as 0.016 microgram / 0.5 ml / mouse. The ELISA titer (0.5097+/-0.0606) was 4 times in the group treated with high dose (0.25 microgram/0.5ml) as in control (0.1113+/-0.0110). Six major protein bands of 66, 55, 35, 33, 18, and 9 kd were detected in Western blot analysis with serum of a vaccine treated mouse, whereas only one weak band of about 35 kd was detected with serum of a control mouse. We concluded that typhoid vaccine produced by purifying Vi antigen of S. typhi very effectively prevent S. typhi infection in mice.
Animals ; Antibodies, Bacterial/immunology ; Antigens, Bacterial/*immunology/*isolation&purification ; Blotting, Western ; Enzyme-Linked Immunosorbent Assay ; Lethal Dose 50 ; Logistic Models ; Male ; Mice ; Mice, Inbred BALB C ; Polysaccharides, Bacterial/*immunology/*isolation&purification ; Salmonella typhi/chemistry/*immunology ; Typhoid Fever/immunology/prevention&control ; Typhoid-Paratyphoid Vaccines/administration & dosage/*immunology/*isolation&purification