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PubMed1998

Bacteraemia following local anaesthetic injections in children.

OBJECTIVES: To estimate odontogenic bacteraemia following three different types of local anaesthetic injections, namely: buccal infiltration analgesia (BIA), conventional intraligamental analgesia (CIA), and modified intraligamental analgesia (MIA). PATIENTS AND METHODS: The bacteraemia-producing potential of three methods of injecting local analgesic solution was determined by taking blood samples, using aseptic technique, from 143 children, aged 1 year 11 months to 19 years 4 months, undergoing general anaesthesia for dental extractions. Of these 143 children, a subgroup of 50 had blood taken before any dentogingival manipulative procedures to provide a baseline level of bacteraemia. The injection methods were buccal infiltration, conventional intraligamental, and a modified intraligamental. The blood samples were taken 30 seconds after injection and cultured in aerobic and anaerobic broth cultures (Bactec) and from lysis filtration vials (Isolator). RESULTS: The percentage prevalence of bacteraemia was: baseline level 8%; buccal infiltration analgesia 16%; modified intraligamental analgesia 50%; and conventional intraligamental analgesia 97%. These values were statistically significantly different using the chi-squared test (P < 0.001). The mean value for colony forming units per millilitre (Isolator system) was 252 (sd = 646) for the intraligamental technique but zero for baseline, infiltration and modified intraligamental techniques. CONCLUSIONS: All local anaesthetic techniques studied were associated with bacteraemia which may have implications for antibiotic prophylaxis for dental treatment. The intraligamental techniques had statistically significantly greater percentage prevalence of bacteraemia compared with baseline. The modified intraligamental technique causes significantly less bacteraemia than the conventional intraligamental technique.

Administration, Buccal↗
PubMed1992

Somatic cell counts in relation to infection status of the goat udder.

Bacteriological analyses, cell counts using the Fossomatic method and California Mastitis Test were performed on 1523 goat milk samples taken aseptically at monthly intervals throughout lactation from three goat herds. Of the goat udders, 81.4% were infected, minor pathogens being the most frequent isolates (65.7%). Differences in the level of infection by minor pathogens were found between herds. Cell counts were influenced by stage of lactation and intramammary infection. Cell counts greater than 10(6) cells/ml were found in 80% of milk samples infected by major pathogens and in 45% infected by minor pathogens. About 81% of udders infected with major pathogens gave California Mastitis Test scores of 2 and 3, compared with 20% for uninfected goats. A high proportion (65%) of udders infected with minor pathogens also produced scores of 2 and 3. A significant positive correlation was found between the California Mastitis Test and cell counts. The use of cell counts for the detection of abnormal goat milk is discussed.

Animals↗
PubMed2002

Bacteriological and epidemiological findings during examination of the uterine content of ewes with retention of fetal membranes.

We included 92 pairs of ewes with or without retention of fetal membranes in a cohort study of 25 flocks in Southern Greece. We obtained two uterine content samples under aseptic conditions, by introducing a swab into the uterus of these ewes, on the 2nd-4th and the 5th-9th day after lambing. We used conventional bacteriological techniques to isolate and identify bacteria and to carry out antimicrobial agents susceptibility testing. The prevalence of bacterial intrauterine contamination among ewes with retention was 24% on the first and 46% on the second sampling (P < 0.0001) and that among ewes without retention was 8 and 2% (P > 0.05), respectively. Clinical signs accompanying the retention of fetal membranes were more frequently observed among ewes with intrauterine contamination than among those without (P = 0.0007). The odds of an ewe having an intrauterine contamination increased multiplicatively by 1.06 when the median duration of retention in the flock increased by 6 h. The principal bacteria isolated from the ewes with retention were Arcanobacterium pyogenes and Escherichia coli; 21% of 73 isolates tested were found resistant to at least one antimicrobial agent.

Actinomycetaceae↗
PubMed2004

Culturable bacteria in subglacial sediments and ice from two Southern Hemisphere glaciers.

Viable prokaryotes have been detected in basal sediments beneath the few Northern Hemisphere glaciers that have been sampled for microbial communities. However, parallel studies have not previously been conducted in the Southern Hemisphere, and subglacial environments in general are a new and underexplored niche for microbes. Unfrozen subglacial sediments and overlying glacier ice samples collected aseptically from the Fox Glacier and Franz Josef Glacier in the Southern Alps of New Zealand now have been shown to harbor viable microbial populations. Total direct counts of 2-7 x 10(6) cells g(-1) dry weight sediment were observed, whereas culturable aerobic heterotrophs ranged from 6-9 x 10(5) colony-forming units g(-1) dry weight. Viable counts in the glacier ice typically were 3-4 orders of magnitude smaller than in sediment. Nitrate-reducing and ferric iron-reducing bacteria were detected in sediment samples from both glaciers, but were few or below detection limits in the ice samples. Nitrogen-fixing bacteria were detected only in the Fox Glacier sediment. Restriction fragment analysis of 16S rDNA amplified from 37 pure cultures of aerobic heterotrophs capable of growth at 4 degrees C yielded 23 distinct groups, of which 11 were identified as beta-Proteobacteria. 16S rDNA sequences from representatives of these 11 groups were analyzed phylogenetically and shown to cluster with bacteria such as Polaromonas vacuolata and Rhodoferax antarcticus, or with clones obtained from permanently cold environments. Chemical analysis of sediment and ice samples revealed a dilute environment for microbial life. Nevertheless, both the sediment samples and one ice sample demonstrated substantial aerobic mineralization of 14C-acetate at 8 degrees C, indicating that sufficient nutrients and viable psychrotolerant microbes were present to support metabolism. Unfrozen subglacial sediments may represent a significant global reservoir of biological activity with the potential to influence glacier meltwater chemistry.

Bacteria↗
PubMed2004

Bladder cell culture on small intestinal submucosa as bioscaffold: experimental study on engineered urothelial grafts.

OBJECTIVES: To investigate the feasibility to perform primary urothelial cell culture using porcine small intestinal submucosa as a delivery scaffold both in vitro and after in vivo implantation in a rabbit model. MATERIALS AND METHODS: Bladder mucosa samples were aseptically obtained from a group of eight male rabbits. The mucosa was cut into fragments and placed on small intestinal submucosa matrices for selective urothelial cell culture. After complete in vitro epithelization the matrices were shaped into tubes and placed in the subcutaneous tissue and subdartos of donor rabbits. The pattern of cell growth and delivery was evaluated on retrieved grafts using histology and immunostaining at the end of the in vitro phase; then 5, 10 and 20 days after implantation. RESULTS: Histological and immunohistochemical analysis of the in vitro primary culture showed the acellular matrices covered with a thin uninterrupted monolayer of urothelial cells. The implants examined on the day 5 maintained the epithelial configuration of the cultured grafts in all samples retrieved. On the day 10 the urothelium showed increased thickness taking on a bilayer configuration. On day 20, all grafts presented the transitional cells arranged in a double layer closely resembling the natural urothelium. The immunostaining pattern displayed the maintaining of urothelial cell phenotype. No differences in epithelium growth and delivery were noted between the two sites of implantation. Five days after implantation, the histological analysis of small intestinal submucosa showed a medium degree tissue reaction with the presence of acute inflammatory cells. Angiogenesis was demonstrated by the development of several new vessels inside the matrix. After twenty days, small intestinal submucosa was gradually replaced with host tissue. CONCLUSION: The small intestinal submucosa proved to function as a means of delivering of autologous urothelial cells cultured in vitro. After ectopic in vivo implantation the bioscaffold maintained viability and growth of the surrounding cells until its degradation.

Animals↗
PubMedDate not supplied

Pathogenicity of different species of staphylococci in caprine udder.

Aseptic foremilk samples were collected from Finnish landrace goats. Ten different species of staphylococci, causing subclinical infections were detected. Twelve goats with persistent subclinical staphylococcal infection were followed on a monthly basis and compared with foremilk samples of nine goats suffering from clinical mastitis. Parameters of inflammation based on the activity of the California Mastitis Test (CMT). N-acetyl-beta-D-glucosaminidase (NAGase) and antitrypsin were determined from the milk. Staphylococci were further classified using the API STAPH system. On the basis of elevations of activities of CMT, NAGase and antitrypsin, Staphylococcus aureus was the most pathogenic in clinical and subclinical mastitis, S. hyicus showed only marginal pathogenicity. Subclinical infections were persistent and the infective organism was not always detected from milk by culture. The biochemical reactions of subclinical staphylococci seemed to vary within the same gland by time. Antitrypsin was most effective in differentiating between subclinical and clinical infection. A teat cistern puncture technique was found to be suitable for the goat.

Animals↗
PubMed2026

Tail-degloving and cold storage facilitate primary cell culture from wild-caught rodents.

The use of wild rodents as biological resources is hindered by the difficulties of live transport and primary culture contamination caused by skin bacteria. To address this, we developed a "tail-degloving" method to physically remove the contaminated external integument. Using the large Japanese field mouse (Apodemus speciosus), this method suppressed contamination compared to conventional biopsies, yielding cells within 5-7 days that were capable of subculturing and cryopreservation. The protocol withstood 4&#xb0;C storage for up to 7 days and enabled contamination-free cell establishment from a large rodent. This tail-degloving protocol eliminates live transport risks and effectively establishes primary cells after a multi-day transport delay.

Aseptic sampling↗
PubMed2001

Staphylococcus aureus associated with mammary glands of cows: genotyping to distinguish different strains among herds.

The hypothesis that strains of Staphylococcus aureus are more likely to be unique to a herd than common to several herds was tested. Herds (n=28) from nine geographic areas of Korea, with elevated milk somatic cell counts (>500000 cells/ml) were enrolled in this study. Mammary quarter milk samples were aseptically collected from all lactating cows (n=616) with at least three functional quarters. Milk was cultured and S. aureus isolates were typed using pulse field gel electrophoresis of DNA SmaI digests. A total of 181 cows were identified as having S. aureus intramammary infections. A total of 52 different types of S. aureus were identified and 34 (65.4%) were associated with a single herd. A total of 18 types of S. aureus were found in multiple herds; 14 types were found in two herds, and four types were found in three herds. Herds with 1, 2, 3, and more than 3 types, were: four (14.3%); eight (28.6%); nine (32.1%); and seven (25.0%). The data indicate that the majority of strains were found in one herd only, and more than 90% were found in two or less herds, suggesting that strains of S. aureus are more likely to be restricted to a single herd, than found in multiple herds.

Animals↗
PubMed2002

Multivariate analysis of determinants of bacterial contamination of whole-blood donations.

BACKGROUND AND OBJECTIVES: Introduction of bacteria into blood components at the collection stage seems to be a frequent occurrence. We therefore assessed determinants of bacterial contamination of whole-blood donations to gain insight into contamination mechanisms and direct prevention. MATERIALS AND METHODS: A cross-sectional study was carried out on donors accepted for whole-blood donation in four French blood banks. Each blood bank used its own two-stage procedure for phlebotomy site preparation. Contamination was identified by culturing two 15-ml samples (collected aseptically at the outset of donation) in a BacT/Alert 240 system. Determinants were assessed by logistic regression analysis. RESULTS: Bacterial contamination, mainly by skin flora, occurred in 76 (2.2%) out of 3385 donations. Significant determinants were as follows: the blood bank (odds ratio [OR] range = 3.0-5.6, P < 0.001); lack of repetition of scrub (OR = 2.7, P = 0.032); and donor age > 35 years (OR = 1.8, P = 0.036). CONCLUSION: Systematic scrub repetition should be implemented to reduce bacterial contamination by skin flora at the collection stage. Further research is required to clarify the role of different antiseptic agents and of donor age.

Adult↗
PubMed1995

Notes on the early history of infective endocarditis and the development of an experimental model.

Experimental models of infective endocarditis antedate Garrison and Freedman's work in 1970. The hypothesis of the role of parasites (microorganisms) microscopically observed in vegetations and cardiac valves of patients with endocarditis was first put forth by Winge in Sweden in 1869. Winge's work led Klebs and Rosenbach in Germany to establish, between 1878 and 1881, an animal model of experimental endocarditis in which the aortic valves of rabbits were perforated with a metallic probe (loaded with septic material) introduced through the carotid artery. Ten years after Winge's work, Pasteur emphasized the importance of bacteriologic "blood cultures." During the period 1881-1886, Netter and Grancher (Pasteur's associates) introduced a method for drawing aseptic blood samples from patients with clinical endocarditis and performing bacteriologic blood cultures. In Vienna in 1885-1886, Orth, Weichselbaum, and Wyssokowitsch further developed Rosenbach's procedure of inducing experimental endocarditis by injecting material from a bacterial culture into a rabbit's ear vein. The development of an experimental model of endocarditis by investigators in the latter part of the nineteenth century provided anatomopathological and bacteriologic data that in turn led to a better understanding of infective endocarditis.

Animals↗
PubMed2003

Identification of spirochetes (treponemes) in endodontic infections.

The purpose of this study was to determine the prevalence of spirochetes in asymptomatic infected root canals and in endodontic abscesses/cellulitis. Aseptic clinical samples were collected using paper points from 54 infected root canals and from aspirates of 84 abscesses/cellulitis. Oligonucleotide primers were produced for PCR identification of Treponema vincentii, T. pectinovorum, T. medium, T. amylovorum, T. denticola, T. maltophilum, and T. socranskii. PCR detected spirochetes in 51 of 84 (60.7%) samples from abscesses/cellulitis and in 20 of 54 (37.0%) samples from asymptomatic infected root canals. T. socranskii was the most frequently detected (44.9%), followed by T. maltophilum (29.7%), T. denticola (28.9%), T. pectinovorum (13.7%), and T. vincentii (5.1%). The number of treponema species detected ranged from 1 to 5 species per sample. The mean numbers of species detected were 2.3 in abscesses/cellulitis and 2.6 in infected root canals. Significant association among species was found between T. maltophilum and T. socranskii, as well as between T. maltophilum and T. denticola by determining the odds ratio (> 2.0).

Bacterial Typing Techniques↗
PubMed2004

Study of bacterial translocation from gut after paraplegia caused by spinal cord injury in rats.

STUDY DESIGN: Normal rats and paraplegic rats with and injured spinal cord were used to study bacterial translocation from gut. OBJECTIVE: To investigate whether bacterial translocation from gut occurs after spinal cord injury. SUMMARY OF BACKGROUND DATA: It has been demonstrated that trauma and operation can lead to gastrointestinal paralysis; disturbance of gastrointestinal motility following trauma may cause bacterial overgrowth in gastrointestinal tract and increase the incidence of bacterial translocation. However, bacterial translocation from gut after spinal cord injury has not been studied. METHODS: Under aseptic manipulation, samples of blood were collected for bacterial cultures and endotoxin determination. In the meantime, samples of mesenteric lymph node, spleen, and liver were collected for bacterial culture. The jejunum and ileum were observed by light and electron microscope. RESULTS: Endotoxemia and bacterial translocation appeared 24 hours and 48 hours correspondingly after spinal cord injury complicated with paraplegia. CONCLUSION: Bacterial translocation from gut would occur after spinal cord injury in rats, which indicated that antibiotics should be administered to paraplegic patients with spinal cord injury as soon as possible to prevent potential bacterial translocation.

Animals↗
PubMed1998

Infectious bovine mastitis caused by environmental organisms.

The purpose of this study was to determine the prevalence of environmental mastitis in dairy herds and identify the main environmental pathogens, and to evaluate the influence of season, housing and management. A total of 20,310 quarters of 5216 animals from 52 dairy herds in 32 counties was examined. Milk samples were aseptically collected for laboratory examination from mammary glands testing positive to any of the field tests. From these, 736 environmental infections were identified. The most frequently isolated environmental pathogens were algae of the genus Prototheca sp. (41.2%), Streptococcus uberis (21.1%), fungi (19.5%), enterobacteriacea (8.3%) and Nocardia sp. (6.6%). The occurrence of mastitis was not influenced by the herd size, use of dry cow therapy, or post milking teat dipping. A tendency for increased occurrence of environmental mastitis during the months of September to February (hot and wet weather) was observed, suggesting a seasonal influence.

Animals↗
PubMed1983

Reduced sulfur in ashes and slags from the gasification of coals: availability for chemical and microbial oxidation.

This study was initiated to determine if reduced sulfur contained in coal gasifier ash and slag was available for microbial and chemical oxidation because eventual large-quantity landfill disposal of these solid wastes is expected. Continuous application of distilled water to a column containing a high-sulfur-content (4% [wt/wt]) gasifier slag yielded leachates with high sulfate levels (1,300 mg of sulfate liter) and low pH values (4.2). At the end of the experiment, a three-tube most-probable-number analysis indicated that the waste contained 1.3 x 10 thiosulfate-oxidizing bacteria per g. Slag samples obtained aseptically from the column produced sulfate under both abiotic and biotic conditions when incubated in a mineral nutrient solution. Both microbial and chemical sulfate syntheses were greatly stimulated by the addition of thiosulfate to the slag-mineral nutrient solution. These results led to a test of microbial versus chemical sulfur oxidation in ashes and slags from five gasification processes. Sulfate production was measured in sterile (autoclaved) and nonsterile suspensions of the solid wastes in a mineral nutrient solution. These ashes and slags varied in sulfur content from 0.3 to 4.0% (wt/wt). Four of these wastes demonstrated both chemical (2.0 to 27 mug of sulfate g day) and microbial (3.1 to 114 mug of sulfate g day) sulfur oxidation. Obvious relationships between sulfur oxidation rate and either sulfur content or particle size distribution of the wastes were not immediately evident. We conclude that the sulfur contained in all but one waste is available for oxidation to sulfuric acid and that microorganisms play a partial role in this process.

Journal Article↗
PubMed2001

Neonatal meningitis in England and Wales: 10 years on.

OBJECTIVES: To determine the incidence of neonatal meningitis in England and Wales. DESIGN: A national postal survey using the British Paediatric Surveillance Unit (BPSU) card scheme supplemented by information from other sources. SETTING: England and Wales 1996-1997. SUBJECTS: A total of 274 babies less than 28 days of age who were treated for meningitis. RESULTS: The incidence of neonatal meningitis in England and Wales has not changed since our previous study in 1985-1987. However, the acute phase mortality has fallen from 19.8% in 1985-1987 to 6.6% in this study. Group B streptococci (42%) and Escherichia coli (16%) remain the most common infecting microorganisms. Eight of 69 (12%) babies with group B streptococci and 4/26 (15%) with E coli died. Antibiotic regimens based on the third generation cephalosporins, notably cefotaxime, were most commonly used (84%). The BPSU scheme identified 72% of cases during the study period. Most cases of viral meningitis were not reported through the BPSU. Less than a third of samples from aseptic meningitis were examined for viruses; 56% of these were positive. CONCLUSIONS: Although the incidence of neonatal meningitis remains unchanged, mortality from this infection has fallen significantly. If this improvement is maintained as reflected in the level of sequelae at 5 years of age, then the fear surrounding meningitis during the neonatal period will have been dramatically reduced.

Anti-Infective Agents↗
PubMed1995

Rate of growth of Pseudomonas fluorescens in donated blood.

AIMS: To examine how delayed refrigeration of blood affects the growth of Pseudomonas fluorescens, one of the two most important causes of sepsis resulting from transfusion of contaminated blood. METHODS: Two donations of whole blood were each divided into three aliquots and inoculated with 5-10 cfu/ml of a P fluorescens strain from a case of transfusion associated sepsis. From each donation, one aliquot was placed at 4 degrees C, one was held at 20 degrees C for six hours prior to refrigeration and the third was held at 20 degrees C for 24 hours prior to refrigeration. Samples were aseptically withdrawn over 17 days and bacterial counts were determined using a pour plate technique. RESULTS: The rate of growth of P fluorescens in blood at 20 degrees C was increased compared with blood at 4 degrees C. At 24 hours the aliquots held at 20 degrees C for six and 24 hours had, respectively, 174 and 29,000 cfu/ml compared with 15 cfu/ml in aliquots held at 4 degrees C. There was no evidence of increased killing of P fluorescens at the higher temperature. CONCLUSIONS: These results suggest that blood for transfusion should be refrigerated as soon as possible after collection.

Blood↗
PubMed2003

Efficacy of intramammary tilmicosin and risk factors for cure of Staphylococcus aureus infection in the dry period.

The objective ofthis study was to evaluate the efficacy of intramammary tilmicosin, administered at drying-off, for eliminating Staphylococcus aureus infection, and to identify risk factors for S. aureus cure during the dry period. A total of 219 naturally infected cows, representing 308 quarters, were randomized to receive either one of two treatments at drying-off. Cows received either an intramammary infusion of 500 mg of benzathine cloxacillin, or a sterile solution containing 1,500 mg of tilmicosin. All cows had quarter milk samples taken aseptically three times before dry-off, and at wk 1, 2, and 4 of the subsequent lactation. Overall, 62% of cows and 67.5% of quarters infected with S. aureus cured during the dry period. The cure following administraton of tilmicosin was 67.3 and 72.5% for cows and quarters, respectively. By comparison, the cure achieved with cloxacillin was 56.9 and 62.9% of cows and quarters. Cows receiving tilmicosin were 2.1 times more likely to cure. The cure rate for cows decreased as the linear score on the last DHI test increased, and as the amount of S. aureus being shed increased. Quarters that cultured positive multiple times before drying-off were less likely to cure. Staphylococcus aureus infections located in front quarters of the udder were 2 times more likely to cure than those in hind quarters. Results of this study demonstrate that intramammary tilmicosin at drying-off is efficacious in curing existing S. aureus during the dry period. Risk factors associated with the cure of S. aureus were identified.

Animals↗
PubMed1983

Effect of parity, stage of lactation, and intramammary infection on concentration of somatic cells and cytoplasmic particles in goat milk.

Aseptic foremilk samples (about 15 ml) were collected biweekly for four samplings and then monthly throughout lactation from 35 does in three herds on Dairy Herd Improvement test. Bacteriology, direct microscopic somatic cell counts, and cytospin differential cell counts were performed on all samples. Milk yield was obtained from monthly Dairy Herd Improvement records. Thirty-nine percent of udder halves were infected, and coagulase negative staphylococci were the most frequent isolates (201 of 205). Somatic cell counts were influenced by lactation number, stage of lactation, and intramammary infection. Also, infection of one udder half increased somatic cells in the corresponding uninfected mammary half of the same animal. Cytoplasmic particles were not affected by stage of lactation or by intramammary infection but were greater for goats in first lactation. Milk yield was influenced by lactation number, stage of lactation, and intramammary infection.

Animals↗