Direct detection of Vibrio cholerae in stool samples

A direct method to detect Vibrio cholerae in stool samples was developed by using a PCR procedure that did not require a DNA purification step. Dilution (1/100) of stool samples prevented inhibition of the reaction by contaminants, and two consecutive PCRs, the second one with a nested primer, achieved the desired sensitivity. Comparison of the results obtained from stool swab samples processed by the two-step PCR and by an enzyme-linked immunosorbent assay using GM1 as the capture molecule showed that the former is more sensitive and gave positive results even when V. cholerae was not culturable or dead.     

H-NS regulates DNA repair in Shigella

We report a new role for H-NS in Shigella spp.: suppression of repair of DNA damage after UV irradiation. H-NS-mediated suppression of virulence gene expression is thermoregulated in Shigella, being functional at 30 degrees C and nonfunctional at 37 to 40 degrees C. We find that H-NS-mediated suppression of DNA repair after UV irradiation is also thermoregulated. Thus, Shigella flexneri M90T, incubated at 37 or 40 degrees C postirradiation, shows up to 30-fold higher survival than when incubated at 30 degrees C postirradiation. The hns mutants BS189 and BS208, both of which lack functional H-NS, show a high rate of survival (no repression) whether incubated at 30 or 40 degrees C postirradiation. Suppression of DNA repair by H-NS is not mediated through genes on the invasion plasmid of S. flexneri M90T, since BS176, cured of plasmid, behaves identically to the parental M90T. Thus, in Shigella the nonfunctionality of H-NS permits enhanced DNA repair at temperatures encountered in the human host. However, pathogenic Escherichia coli strains (enteroinvasive and enterohemorrhagic E. coli) show low survival whether incubated at 30 or 40 degrees C postirradiation. E. coli K-12 shows markedly different behavior; high survival postirradiation at both 30 and 40 degrees C. These K-12 strains were originally selected from E. coli organisms subjected to both UV and X irradiation. Therefore, our data suggest that repair processes, extensively described for laboratory strains of E. coli, require experimental verification in pathogenic strains which were not adapted to irradiation.     

Shigellosis in children: a clinico-epidemiological comparison between Shigella dysenteriae type I and Shigella flexneri

A new method was developed to measure the content of a Lumbricus component in a traditional Chinese medicine (TCM). An antiserum specific to Lumbricus was elicited in a rabbit following immunization with a suspension of Lumbricus fragments. A characteristic antigen protein, 70 kDa, was found in Lumbricus and was purified almost to singleness using a column chromatography series of gel filtration and DEAE-Sepharose. A selected antibody enzyme immunoassay (SAEIA) was developed using the antiserum and the purified 70 kDa protein as a solid-phase antigen. The SAEIA was specific to Lumbricus species, and showed no cross-reaction with any crude drugs other than Lumbricus. This SAEIA detected 70 kDa protein in the amount of 10 ng/ml with excellent reproducibility (coefficient of variation=3.0%) and an EC50 of 0.24 microg/ml. Using this assay, Lumbricus levels were easily determined in a Lumbricus-based TCM Kazecoll, but not in the control Kazecoll (Kakkonto) prepared without Lumbricus. The SAEIA for 70 kDa protein was simple, accurate, reproducible and may provide a general analytical method for the quality control of Lumbricus-based TCMs.     

Screening of stool samples for identification of vancomycin-resistant Enterococcus isolates should include the methyl-alpha-D-glucopyranoside test to differentiate nonmotile Enterococcus gallinarum from E. faecium

The methyl-alpha-D-glucopyranoside (MDG) test has been shown to be superior to motility testing in differentiating Enterococcus faecium from E. gallinarum. In the present study, 33 vancomycin-resistant enterococcus (VRE) isolates collected as part of a stool surveillance study were compared by using motility and MDG. Motility testing identified all 33 isolates as E. faecium, whereas MDG identified 11 of the 33 isolates as nonmotile E. gallinarum. The MDG results were confirmed by sequencing the 16S rDNA V6-to-V8 region. We conclude that the MDG test is a necessary component of routine VRE screening.     

Shigella and the fluorinated quinolones

An 8-year-old girl presented with marked shortening of the right forearm due to destruction of both the radius and ulna secondary to neonatal osteomyelitis. A one-bone forearm operation was performed to achieve a stable forearm. Two years later, the one-bone forearm was lengthened for 6 months by callus distraction (callotasis) achieving 12 cm of extra length. The patient was last followed up at the age of 16. The appearance and functional outcome of the right upper limb had been improved and she was independent in all activities of daily living.     

Times of first void and first stool in 500 newborns

The times of the first stool and the first void were recorded for 500 infants. The population studied was defined in terms of gestational age, maternal premedication, and mode of delivery. The results are presented in tabular form by time segments with term, preterm, and postterm infants treated separately.     

New provisional serovar of Shigella boydii

Ten bacterial strains are described that give biochemical reactions characteristic of the genus Shigella but do not belong to any of the established or provisional serovars. The strains ferment mannitol and are antigenically indistinguishable. They do not possess any of the group antigens of Shigella flexneri, and it is therefore proposed that they be regarded as members of a new provisional serovar of Shigella boydii. Strain E16553 is designated as the test strain for the new serovar.     

Mechanism of bacteriophage SfII-mediated serotype conversion in Shigella flexneri

We have isolated the lysogenic bacteriophage SfII, which mediates glucosylation of Shigella flexneri O-antigen, resulting in expression of the type II antigen. SfII belongs to group A of the Bradley classification and has a genome size of 42.3kb. DNA sequencing of a 4 kb BamHI subclone identified four open reading frames (ORFs), of which only two were found to be necessary for serotype conversion. These genes were named bgt, which encodes a putative bactoprenol glucosyl transferase, and gtrII, encoding the putative type II antigen determining glucosyl transferase. These genes are adjacent to the integrase gene (int) and attachment site (attP), which are highly homologous to those of Salmonella bacteriophage P22. Another ORF encoded a highly hydrophobic protein of 120 amino acids with homologues in Escherichia coli, Salmonella bacteriophage P22 and S. flexneri. Previous studies identified gtrX, the glucosyl transferase gene, of bacteriophage SfX, which also glucosylates the O-antigen specifically. We determined that gtrX-mediated expression of the group 7,8 antigen also requires bgt. This allowed us to identify gtrII as being the serotype antigen II determining glucosyl transferase. Southern hybridization and polymerase chain reaction (PCR) analyses indicated that bgt homologues exist in the genomes of all S. flexneri serotypes and in E. coli K-12, whereas gtrII was only detected in strains of serotype 2. Transposon TnphoA-derived chromosomal mutations of bgt and gtrII in S. flexneri serotype 2a were isolated and characterized. [35S]-methionine labelling and the use of a T7 RNA polymerase expression system identified a protein of 34kDa corresponding to Bgt. However, GtrII, which has a predicted molecular weight of 55 kDa, was not detected. We propose that the function of Bgt is to transfer the glucose residues from the UDP-glucose onto bactoprenol and GtrII then transfers the glucose onto the O-antigen repeat unit at the rhamnose III position. The chromosomal organization of these serotype-converting genes, when compared with their homologues in E. coli K-12 chromosome and the P22 bacteriophage genome, were very similar. This suggests that the regions encode similar functions in these organisms and have a similar evolutionary origin.     

Resistance to antibiotics of Shigella strains isolated in Somalia

The resistance to antibiotics of 240 Shigella strains isolated in Somalia from 1973 to 1976 was studied. Many strains, particularly those of Shigella dysenteriae type 1, were found to be resistant to more than one drug. In view of their resistance to ampicillin, chloramphenicol, streptomycin, tetracycline, and sulfonamides, it is suggested that polymyxin B or M sulfate - which have proved to be effective in vivo - should be used for the treatment of clinically typical cases of bacillary dysentery.     

FECOM: a new artificial stool for evaluating defecation

OBJECTIVE: Traditionally, barium paste has been used for performing defecography. Because this substance is not stool-like, barium defecography may not accurately represent defecatory function. Our aim was to prospectively compare the utility of a new artificial stool, "FECOM"--a silicon-filled and barium-coated, deformable device the shape and consistency of which mimicked a normal formed stool with that of barium paste. METHODS: Defecography was performed after placing FECOM or barium paste in a random order in 12 healthy subjects (two men and 10 women). We evaluated the changes in anorectal angle, rectal morphology, rectal sensation, and the subjects' preference for a "stool-like" device. RESULTS: Anorectal angle at rest, during squeeze, cough, and straining were each greater with the FECOM when compared with the barium paste (p < 0.006). Anterior rectocele (nine), mucosal intussusception (four), and incontinence (three) were identified only with barium defecography. Nine (75%) subjects preferred FECOM to barium paste (p < 0.001) and reported that expulsion of this device mimicked more closely their stools at home (p < 0.05). CONCLUSION: The anorectal angle is influenced by the form and consistency of stool material and is lower with barium paste. The detection of rectocele, mucosal intussusception, and barium leakage in normal subjects during barium defecography questions the significance of these findings. FECOM appears to be a realistic alternative to barium paste for performing defecography.     

771 Shigella strains isolated in a 15-year period from Americans living in Ankara. Comparisons with Shigella strains isolated from Turkish natives

The author had worked as a bacteriologist in Ankara American Air Force Hospital from 1958 to 1972. During this period of time he was able to isolate 771 shigella strains from the American patients of the hospital. The amounts and the percentages of the subgroups are listed below together with the two other Ankara City Pediatric hospital results (see article). american community lived in Ankara for 15 years and shared many aspects which were important in shigella epidemiology like using city's tap water, shopping from the same green groceries to by fruits and vegetables so on. For shigella cases Ankara had seasonal alterations in July and August being the peak months. Americans, disease followed that course exactly. The big difference was being the local people of the city had those epidemics with the flexneri strains but Americans had them with the sonnei strains. To show the possible sources which could infect the community time to time and make sonnei predominant yielded negative result. All the patients stools were cultured. Sensitivity studies employing "plate disk diffusion" techniques were done. All patients treated accordingly. Stools checked for routine carriership exams. None become a chronic carrier. American installations had their shops, clubs, mess halls. The food handlers of these places had checked routinely by the preventive medicine office. None found infected. Only at 1963 the amount of flexneri isolations were higher than sonnei. All the remaining years sonnei were predominant. Antibiotic sensitivity studies showed no differences between subtypes. Except streptomycin and sulphamide strains stayed fairly sensitive to other antibiotics. Resistance factors contained maximum 6 markers. The little Americans community in Ankara consisted a unique and highly socioeconomic-wise developed group of the city. Many attempts to find the answer for the question is "who is infecting who" was always fruitless. That gives the impression as if the Americans were more sensitive to the shigella infections caused by sonnei strains for some reason. In another words Americans were more resistant to the infections other than sonnei.     

Heterophyid eggs in human stool samples in Assam: first report for India

Two cases from a Muslim family in upper Assam were found to pass trematode eggs morphologically similar to the members of the family Heterophyidae Odhner, 1914. The eggs characteristically yellowish brown in colour, ovoid, have a thick shell and are operculate. They measured 31.5 +/- 2.0 um SD x 22.4 +/- 1.9 um SD with well developed miracidium. This is the first record of human heterophyid infection from India.     

Detection of decay-accelerating factor in stool specimens of patients with colorectal cancer

BACKGROUND & AIMS: Colorectal cancers have an increased expression of decay-accelerating factor (DAF). The aim of this study was to determine whether stool specimens of patients with colorectal cancer contain increased amounts of DAF. METHODS: DAF was measured using an immunoassay in the stool specimens of 40 persons with colorectal cancer, 18 with colorectal adenomatous polyps, 13 with upper gastrointestinal cancer, and 41 without gastrointestinal disease. RESULTS: Stool DAF concentrations in patients with colorectal cancer (0-9.8 ng/g stool; median, 1.6 ng/g) were significantly higher than those in patients with adenoma (0-6.4 ng/g; median, 0 ng/g) (P < 0.05), patients with upper gastrointestinal cancer (0-3.1 ng/g; median, 0 ng/g) (P < 0.05), and subjects without gastrointestinal disease (0-3.4 ng/g; median, 0 ng/g) (P < 0.01). Resection of colorectal cancers caused a marked decrease in stool DAF concentrations. The stool DAF test was positive in a substantial portion of patients with colorectal cancer whose tumors were small ( < 2 cm), at an early TNM stage, or unassociated with fecal occult blood positivity. The sensitivity of the test for colorectal cancer was 55%, and the specificity was 85%. CONCLUSIONS: The measurement of stood DAF deserves evaluation as a test for detection of colorectal cancer.     

Strategy for cross-protection among Shigella flexneri serotypes

Based upon the lipopolysaccharide (LPS) structure and antigenicity of Shigella group B, a strategy for broad cross-protection against 14 Shigella flexneri serotypes was designed. This strategy involves the use of two S. flexneri serotypes (2a and 3a), which together bear the all of the major antigenic group factors of this group. The novel attenuated strains used in these studies were S. flexneri 2a strain CVD 1207 (DeltaguaB-A DeltavirG Deltaset1 Deltasen) and S. flexneri 3a strain CVD 1211 (DeltaguaB-A DeltavirG Deltasen). Guinea pigs were immunized with an equal mixture of these strains and later challenged (Sereny test) with a wild-type S. flexneri serotype 1a, 1b, 2b, 4b, 5b, Y, or 6 strain of demonstrated virulence in the same model. Guinea pigs that were immunized with these two vaccine strains produced serum and mucosal antibodies that cross-reacted with all the S. flexneri serotypes tested (except of S. flexneri serotype 6) as assessed by enzyme-linked immunosorbent assay, immunoblotting, and slide agglutination. Furthermore, the combination vaccine conferred significant protection against challenge with S. flexneri serotypes 1b, 2b, 5b, and Y but not with serotypes 1a, 4b, or (as predicted) 6.     

Antimicrobial susceptibility of Shigella from a rural community in Bangladesh

Of the 63 Shigella strains isolated from stool cultures from 200 patients who attended a district hospital in Bangladesh with bloody diarrhoea, 37 (59%) were S. dysenteriae type 1, 25 (39%) were S. flexneri and only one (2%) was S. sonnei. Over half (54%) of the Shigella isolates came from children aged < 10 years. Most (89%) of the isolates of S. dysenteriae type 1 were resistant to ampicillin, cotrimoxazole, nalidixic acid, tetracycline and chloramphenicol. Although many (60%) of the isolates of S. flexneri were resistant to ampicillin and cotrimoxazole, only 4% of them were resistant to nalidixic acid. However, all of the S. dysenteriae and S. flexneri were sensitive to ciprofloxacin. The need for periodic monitoring to determine the resistance pattern in remote areas is emphasised.     

Identification of Shigella flexneri subserotype 1c in rural Egypt

In a population-based study of diarrhea in rural, northern Egypt, 60 Shigella flexneri strains were identified, of which 10 could not be definitively serotyped. Serological analysis with commercial reagents suggested that they were serotype 1, but the strains failed to react with subserotype 1a- or 1b-specific antibodies. All 10 strains reacted with MASF 1c, a monoclonal antibody specific for a provisional S. flexneri subserotype, 1c, first identified in Bangladesh and not previously detected outside of that region. Our results show that S. flexneri subserotype 1c is not unique to Bangladesh and that the inability to detect it may reflect both the limited use of suitable screening methods and the rarity of this subserotype.