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R Möllby

Publications and source records attributed to R Möllby.

At least 55 records · Page 3Linked to original sources

Phenotypic characterization of intestinal Escherichia coli of pigs during suckling, postweaning, and fattening periods.

A highly discriminatory and standardized biochemical fingerprinting method was used to monitor the persistence and colonization of intestinal Escherichia coli isolated from the feces of four sows and their litters (four piglets from each) during the suckling, postweaning, and fattening periods. Altogether, 195 fecal samples were collected and 1,827 E. coli strains were tested (mean number of isolates tested per fecal sample per pig, 9.5). Strains were divided into similarity groups on the basis of their biochemical phenotypes (BPTs). The diversity of E. coli strains in each sample was measured with Simpson's index of diversity, and similarity between E. coli floras of piglets was calculated with a population similarity index. Each fecal sample contained several BPTs of E. coli, some of which dominated that population. The intestinal colonization of piglets consisted of successive waves of different E. coli BPTs, the tenure of which varied from a few days to 2 weeks. Most of these BPTs disappeared in the succeeding samples and were not recovered again from the same piglets. On the other hand, some E. coli strains which colonized piglets early during the suckling period persisted for a long period and were referred to as resident BPTs. Each piglet carried more than one resident BPT (mean of 2.4 BPTs per pig), some of which were also found in other piglets.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Glutamine supplementation does not prevent bacterial translocation after non-lethal haemorrhage in rats.

OBJECTIVE: To find out whether supplementation of an enteral diet with glutamine would reduce translocation of bacteria to mesenteric lymph nodes or blood after major haemorrhage in rats. DESIGN: Open randomised study. SETTING: University departments of surgery and microbiology, Sweden. MATERIAL: 49 Sprague-Dawley rats. INTERVENTIONS: Rats were fed enterally for 7 days on diets supplemented with either glutamine or an isonitrogenous amount of non-essential amino acids. After feeding, 8 experimental and 8 control rats underwent sham operation; 9 and 7, respectively, underwent moderate haemorrhage (to 65 mm Hg); and 9 and 8, respectively, underwent severe haemorrhage (50 mm Hg) without reinfusion. MAIN OUTCOME MEASURES: Microbiological analyses of samples of blood and mesenteric lymph nodes taken 24 hours after haemorrhage. RESULTS: The median (interquartile) number of colony forming units/mesenteric lymph nodes after moderate haemorrhage in animals who were given glutamine supplementation was 11 (0-34) and in control animals 20 (0-178). After severe haemorrhage the corresponding figures were 199 (10-310) and 22 (0-187). No pathogens were isolated from blood cultures. CONCLUSION: Glutamine supplementation before haemorrhage did not reduce bacterial translocation to mesenteric lymph nodes in this rat model.

Animals↗

The Gal(alpha 1-4)Gal-specific tip adhesin of Escherichia coli P-fimbriae is needed for pyelonephritis to occur in the normal urinary tract.

Nonobstructive acute pyelonephritis in humans is most often caused by P-fimbriated Escherichia coli. P-fimbriae are heteropolymeric fibers carrying a Gal(alpha 1-4)Gal-specific PapG adhesin at its distal end. The pyelonephritic strain DS17 expresses P-fimbriae from a single gene cluster. A mutant strain, DS17-8, which expresses P-fimbriae tacking the PapG adhesin, was constructed by allelic replacement introducing a 1-bp deletion early in the papG gene. In cynomolgus monkeys, DS17 and DS17-8 were equally able to cause bladder infection, whereas only the wild-type strain DS17 could cause pyelonephritis as monitored by bacteriological, functional, and histopathological criteria. Since DS17, but not DS17-8, adheres to renal tissue, these data underscore the critical role of microbial adherence to host tissues in infectious disease and strongly suggest that the PapG tip adhesin of P-fimbriae is essential in the pathogenesis of human kidney infection.

Adhesins, Escherichia coli↗

Composition and diversity of intestinal coliform flora influence bacterial translocation in rats after hemorrhagic stress.

Coliform bacteria are the most frequently reported bacteria to translocate after hemorrhage. We investigated the correlation between composition and diversity of the cecal coliform flora and the degree of translocation in a rat model of hemorrhagic stress. Two groups of nine rats each were bled to 60 and 50 mm Hg mean arterial blood pressure, respectively. A sham-operated group without bleeding (n = 9) and a noninstrumented group (n = 6) served as controls. From each rat, 40 coliform isolates from the cecum and up to 16 from positive mesenteric lymph node (MLN) cultures were tested with an automated biochemical fingerprinting method. The phenotypic diversity of coliforms in each cecal sample was calculated as Simpson's diversity index (DI), and similarities between bacterial types in different samples were calculated as population similarity coefficients. Three rats in the sham-operated group and seven in each of the bled groups showed bacterial translocation. Of the different biochemical phenotypes (BPTs) found in the cecum of bled rats (mean, 6.5 BPTs), only a few were detected in MLNs (mean, 1.9 BPTs per MLN), with Escherichia coli being the dominant species. The translocating E. coli strains were mainly of two BPTs. Rats showing no translocation either did not carry these strains or had a high diversity of coliforms in the cecum. Furthermore, translocation of these coliform types was independent of their proportion in the cecum. In bled rats, the diversity of coliforms (mean DI, 0.53) was significantly higher than that in control groups (mean DI, 0.30; P = 0.004), suggesting that hemorrhage stimulates an increase in diversity of cecal coliforms. Rats with similar coliform flora and subjected to the same treatment showed similar patterns of translocation. Our results suggest that the composition of the coliform flora is an important factor in translocation and that certain coliform strains have the ability to translocate and survive in MLNs more easily than others.

Animals↗

Biochemical phenotypes of Salmonella Livingstone isolated from humans, animals and feedstuffs in Sweden.

Salmonella Livingstone is occasionally isolated from humans, animals and feedstuffs in Sweden. To follow the spread of infection and trace the source of isolates, adequate typing methods are needed. We have developed an automated typing system based on biochemical fingerprinting of bacteria (the PhP system) for typing of different Salmonella serotypes. The system measures the kinetics of various biochemical reactions of bacteria grown in liquid medium in microtiter plates and uses numerical techniques to identify biochemical phenotypes (BPTs) among the tested strains. In the present study we used a set of 16 highly discriminatory tests to differentiate strains of Salmonella of serotype Livingstone and evaluated the system for its discriminatory ability using a collection of 34 unrelated human isolates of S. Livingstone. We also used the system to investigate BPTs of 45 Livingstone strains isolated from animals and feedstuffs in Sweden between 1987 and 1991. Altogether 19 different BPTs were found among human isolate giving a diversity index (Di) of 0.930. In contrast, most strains isolated from animals and feedstuffs in Sweden belonged to 2 dominating BPTs (Di = 0.704). One of these contained 17 strains mainly isolated during 1992 whereas the other contained 18 strains isolated between 1987 and 1991. None of the Swedish human isolates were identical to those of animals and feedstuffs. These findings suggest that 2 different BPTs of Salmonella Livingstone strains are particularly common among animals and feedstuffs in Sweden and that they are not related to human cases of enteritis in this country. We also conclude that biochemical fingerprinting with the PhP system is a reliable and highly discriminatory method for detecting epidemic strains of Salmonella Livingstone.

Animal Feed↗

Pathogenesis of urinary tract infection--experimental studies of vaginal resistance to colonization.

The review summarizes studies of vaginal colonization resistance against Escherichia coli in a primate model. The genital flora surrounding the urethral orifice exerts a strong colonization resistance. Amoxicillin profoundly disturbs the normal vaginal microflora, reduces its adherence to vaginal epithelial cells in vivo and promotes a persistent vaginal E. coli colonization. Certain cephalosporins may have a similar effect. The induced ecological changes mimic those seen in patients with recurrent urinary tract infections (UTI). Amoxicillin also promotes the spread of E. coli from rectum to vagina, which may be of clinical significance. Trimethoprim and nitrofurantoin do not have these effects. The natural colonization resistance could not clearly be correlated with the presence of lactobacilli, which were only transiently reduced by amoxicillin. The colonization resistance against E. coli could only partly be restored by vaginal instillation of lactobacilli, but was fully restored by flushing of the whole vaginal flora from a healthy monkey. Clinical observations suggest that accumulation of E. coli around the urethral orifice increases the risk of UTI. We conclude that antibiotics and other compounds that interfere with the normal genital flora may increase the risk of UTI. This should influence the choice of antibiotics in the treatment of UTI.

Animals↗

Diagnostic evaluation of polymerase chain reaction discriminative for Bordetella pertussis, B. parapertussis, and B. bronchiseptica.

A polymerase chain reaction (PCR) procedure for simultaneous detection and identification of Bordetella pertussis, B. parapertussis, and B. bronchiseptica was developed and evaluated against culture in a study comprising nasopharyngeal aspirates and swabs from 166 patients with suspected pertussis, 54 of which were culture positive. A 239-base-pair sequence in the pertussis toxin promoter region was amplified using primers Bouni 1: 5'GCACCATCCCGCATACGTGTTG3', and Bouni 2: 5'GTGCAACGCATCCCGTCTTCC3'. The sequence contains mutations in B. parapertussis and B. bronchiseptica, and species were differentiated by restriction enzyme cleavage of the amplified product. The lowest detectable amount of B. pertussis DNA was 0.1 pg (equals approximately 30 bacteria). No false positives were found in clinical samples or among 18 other species. Treatment of 66 aspirates with a weak cation exchange resin increased the diagnostic sensitivity of PCR. Two culture-positive aspirates were negative by PCR, but grew with a single colony among contaminating flora and could be identified only after PCR analysis of the colony material. The amount of positive cases was increased from 13 by culture to 19 by the addition of PCR. Six samples positive by PCR were culture negative. All six patients showed clinical and epidemiologic evidence of pertussis, and three patients had been treated with antibiotics. PCR increased the sensitivity of pertussis case finding with retained specificity and can be used for laboratory diagnosis of whooping cough.

Base Sequence↗

Variations in biochemical phenotypes and phage types of Salmonella enteritidis in Germany 1980-92.

The Phene Plate system for typing Salmonella serotypes (PhP-S) is a simple automated typing method based on biochemical fingerprinting. It gives a quantitative value of the metabolism of various substrates by measuring the speed and intensity of each reaction. The 'biochemical fingerprint' of each isolate is used to calculate similarities among the tested strains with a personal computer program. We used this system to examine a collection of 86 strains of Salmonella enteritidis isolated from human sporadic cases in Germany between 1980 and 1992. Twenty-three biochemical phenotypes (BPTs) consisting of 9 common (C) and 14 single (S) BPTs were identified. BPTs C2 and C4 containing 20 and 36 strains respectively accounted for 65% of the isolates. Strains of BPT C2 were found over a wide period of time whereas strains of BPT C4 were isolated during the period between 1988 and 1992. With phage typing, 11 discrete phage types (PTs) and 18 strains designated as non-specific type (NST) were identified. PTs 4 and 8 with 39 and 17 strains respectively were the dominant PTs. Strains of PT 8 were isolated over a wide period of time whereas all (except one) strains of PT 4 were isolated between 1988 and 1992. Combination of biochemical fingerprinting and phage typing divided the strains into 25 phenotypes (BPT:PTs). Whilst phenotype C2:8 was found over a number of different years, phenotype C4:4 was isolated only between 1988 and 1992. These findings indicate the presence of one persistent and one recently emerged phenotype among S. enteritidis strains in Germany.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacterial Typing Techniques↗

Evaluation of new enzyme-linked immunosorbent assay based on a supernatant containing Staphylococcus aureus alpha-toxin produced by Bacillus subtilis.

The gene encoding alpha-toxin from Staphylococcus aureus was cloned into a Bacillus subtilis expression vector (pEF 231/alpha-Tox). The protease-deficient B. subtilis strain DB 104 transformed with pEF 231/alpha-Tox expressed and secreted 5 mg of alpha-toxin per liter into the growth medium. The alpha-toxin-containing supernatant was diluted 200-fold and used as coating antigen in an enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of septicemia and endocarditis caused by S. aureus. Paired sera from patients in acute and convalescent stages of S. aureus and non-S. aureus infections were used to evaluate this ELISA. To evaluate the effectiveness of the crude preparation, the results were compared with those of an ELISA based on a commercially available alpha-toxin. Similar rises in serum titers were obtained with either type of alpha-toxin preparation. This is the first time a crude supernatant without any further purification has been used as an ELISA coating antigen. We therefore conclude that B. subtilis is a suitable host organism for cheap and simple production of prokaryotic recombinant antigens to be used in serodiagnosis.

Antibodies, Bacterial↗

A prospective study of first attacks of inflammatory bowel disease and non-relapsing colitis. Microbiologic findings.

In 105 patients with a first attack of colitis, thorough microbiologic investigations of rectal biopsy, faecal, and serum samples were performed with the aims of identifying the colitis-causing agents and shedding light on factors that may precipitate or aggravate the onset of inflammatory bowel disease. Sixty-one patients were found to have inflammatory bowel disease. In 13 (21%) of these patients microbial findings were positive. Eight of the 61 patients fell ill during or immediately after antibiotic treatment, and 10 while travelling abroad. Forty-one of the 105 patients had non-relapsing colitis. In 32 (78%) of these the microbial findings were positive. Six of these 41 patients fell ill during or immediately after antibiotic treatment, and 14 while travelling abroad. Alteration of the intestinal microflora on travelling, gastrointestinal infection, or treatment with antibiotics seems to precipitate or aggravate the symptoms in latent inflammatory bowel disease. In such patients the mode of onset is often changed from insidious to more acute, which may cause difficulty in differentiation from non-relapsing colitis.

Acute Disease↗

Bacterial translocation after non-lethal hemorrhage in the rat.

Translocation of enteric bacteria has been suggested to compromise patients in severe catabolic stress. Mechanisms for this route of infection are not known. In this study, rats were subjected to hemorrhage without reinfusion during 60 min, total blood loss was 3.28 +/- 0.14 ml/100 g BW. Control groups consisted of sham-operated animals without bleeding, and rats not operated at all. The mean number of viable bacteria found in mesenteric lymph nodes (MLN) of bled animals was 168 +/- 45 colony forming units (c.f.u./MLN), significantly higher compared to sham operated (5 +/- 3 c.f.u./MLN) and not operated (0 +/- 0 c.f.u./MLN) controls (P < 0.01). Cultures from MLN were positive in 7/9 rats after bleeding, in 3/9 of sham operated, and in 0/6 of non-instrumented control animals. No positive blood cultures were isolated. Escherichia coli was the dominant species found in MLN. A biochemical fingerprinting method (the PhP system) was used to identify translocating strains of E. coli among strains found in cecum. The method was also used to compare translocating strains between different animals. Our findings reveal that bacteria translocate to MLN after hemorrhage. Some phenotypes of E. coli strains translocate more frequently than others, suggesting that they have properties facilitating translocation.

Animals↗

The importance of P and type 1 fimbriae for the persistence of Escherichia coli in the human gut.

The faecal Escherichia coli flora was studied in 89 infants. Each infant was followed with a mean of 12 faecal samples (range 5-21) between 0 and 18 months of age. All isolates were assayed for P fimbriae and biochemically phenotyped and the persistence of each strain (phenotype) in the infant's gut was determined. In a subset of strains the occurrence of type 1 fimbriae and adherence to HeLa cells was studied. Thirty-one per cent of isolates belonging to strains colonizing for longer than 6 months expressed P fimbriae compared to 19% of the isolates from strains colonizing 1-6 months or transient strains colonizing less than 1 month. Type 1 fimbriae and adherence to HeLa cells occurred similarly often in all groups of strains. We conclude that P fimbriae, but not type 1 fimbriae or HeLa cell adherence seemed to contribute to the ability of the E. coli strain to colonize the human intestine.

Bacterial Adhesion↗

The effect of amoxycillin on vaginal colonization resistance and normal vaginal flora in monkeys.

Previous studies have shown that vaginal colonization resistance in monkeys can be eliminated by amoxycillin and restored by flushing vaginal flora from a healthy monkey into the vagina of a monkey colonized with Escherichia coli. The hypothesis that the effect of amoxycillin resulted from elimination of parts of the normal flora was tested in the present study. Nine monkeys were flushed vaginally with amoxycillin daily for six days. The number of anaerobic bacteria decreased during amoxycillin administration, as did the number of species isolated. The most obvious effects were observed among the genera Bacteroides and Peptostreptococcus, while Lactobacillus spp. were less affected. Restoration of the flora after amoxycillin administration was slow in most of the monkeys. During amoxycillin administration, all monkeys became colonized spontaneously with E. coli. This was not, however, associated with increased adherence in vitro. The colonization persisted throughout the study period (29 days). It was concluded that amoxycillin disturbs vaginal colonization resistance by eliminating at least part of the normal vaginal flora, thereby promoting periurethral colonization with enterobacteria.

Amoxicillin↗

Application of biochemical fingerprinting to the investigation of clonal groups of Salmonella of serotype Havana.

A computerised typing method based on biochemical fingerprinting was used to investigate biochemical phenotypes (BPTs) among 70 strains of Salmonella of serotype Havana isolated from human cases of gastroenteritis in Iran and other parts of the world. A total of 16 BPTs comprising five common and 11 single phenotypes was identified. The most frequently found BPT contained 24 isolates from Iran and nine from other countries. Three common BPTs with two, seven and 15 isolates were found among Iranian strains only and one common BPT with two isolates was found among non-Iranian strains only. Antibiotic-resistance patterns and virulence properties of strains from these common BPTs suggested that they might be unique clones. Forty-two Iranian isolates shared multi-resistance to between three and seven antibiotics. In contrast, none of the isolates from other countries was resistant to antibiotics. Furthermore, 43 Iranian isolates showed mannose-resistant adhesion to HeLa cells and 24 of them possessed an aerobactin-mediated iron-uptake system, whereas none of the isolates from other countries possessed any of these virulence properties. These findings suggest that four unique clones of Salmonella Havana with different BPTs and virulence properties are common in Iran; two particular clones were responsible for a majority of Havana infections there. However, the most prevalent BPT found among Iranian strains was also common in strains from other countries. It is concluded that biochemical fingerprinting, as used in this study, is a reliable method for identifying clonal groups of Havana strains. The method is reproducible, easy to perform and can be used alone, or in combination with other typing methods, in epidemiological studies of serotype Havana.

Bacterial Adhesion↗

Evaluation of the PhP system for biochemical-fingerprint typing of strains of Salmonella of serotype Typhimurium.

The Phene Plate (PhP) system of biochemical fingerprinting of bacteria is a computerised typing system, based on quantitative measurements of the kinetics of several biochemical reactions of bacteria grown in liquid medium in microtitration plates. For each isolate tested, it yields a biochemical fingerprint comprising several kinds of quantitative data which are useful for establishing similarities among strains with a personal-computer program. In this study, a set of 16 specific substrates was chosen to differentiate strains of Salmonella of serotype Typhimurium. The system was evaluated for its typability, reproducibility and discriminatory power in tests with a collection of 100 epidemiologically unrelated Typhimurium strains and results were compared with those obtained by phage typing. At an identity level of 0.980, strains were assigned by this method to 51 biochemical phenotypes (BPTs), giving a diversity index of 0.963 and a resolution index of 0.210. In contrast, 24 phage types (PTs) were identified among these isolates (a diversity index of 0.901). The combined use of biochemical fingerprinting by the PhP system and phage typing discriminated 82 phenotypes (a diversity index of 0.994). Stability of markers in each of the methods was also evaluated after subculture of 20 strains for 21 consecutive days. Only nine biochemical reactions were found that were subject to small, but measurable, changes for at least one isolate. These changes slightly decreased the mean similarity coefficients among strains but the overall BPTs of the strains showed changes in four strains (20%). In contrast, eight strains (40%) showed changes in their PTs after this treatment. It is concluded that the PhP system is a highly discriminatory and reproducible method for typing Typhimurium strains. It is easy to perform, and may be used alone or in combination with phage typing in epidemiological studies of Typhimurium strains.

Bacterial Typing Techniques↗

The use of biochemical fingerprinting, phage typing and antimicrobial-susceptibility testing in the detection of epidemic strains of Salmonella of serotype Typhimurium in Iran.

A collection of 86 strains of Salmonella of serotype Typhimurium isolated from children with gastroenteritis in Tehran, Iran was examined for biochemical phenotype, phage type and antibiotic-resistance pattern. Twenty-seven biochemical phenotypes (BPTs), 14 discrete phage types (PTs) and 18 resistotypes (RTs) were identified. Fifty-three strains (62%) belonged to two major and probably related BPTs, whereas the other 33 isolates belonged to less common BPTs. The two predominant BPTs contained 26 strains of the same PT and 23 strains of the same RT. Different PTs and RTs of strains with similar BPT were sometimes observed, possibly reflecting antibiotic pressures in Iran. These results suggest that two major "clones" of Typhimurium strains are particularly common in Iran and, although each method alone adequately detected these and other less common "clones", biochemical fingerprinting provided additional information about relationships among strains.

Bacterial Typing Techniques↗

Lipase versus teichoic acid and alpha-toxin as antigen in an enzyme immunoassay for serological diagnosis of Staphylococcus aureus infections.

Titres of IgG antibodies to Staphylococcus aureus lipase were analysed in 448 sera from patients suspected of having Staphylococcus aureus infections and the results compared to those for the routinely used staphylococcal antigens teichoic acid and alpha-toxin. The results indicated that determination of serum antibodies to lipase is a sensitive assay for serological diagnosis of staphylococcal infections and increased sensitivity may be achieved by selection of optimal antigen combinations.

Adolescent↗