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[Clinical and laboratory findings in a series of cases of infective endocarditis].

OBJECTIVE: To describe clinical and laboratory data of infective endocarditis (IE) in 28 children from Vitória, state of Espírito Santo, Brazil. METHODS: We reviewed the medical records of 28 children aged 18 years and under admitted to the Infectious Diseases Unit of Nossa Senhora da Glória Children's Hospital with a diagnosis of IE from January 1993 to December 2001. The diagnosis of IE was based on the criteria established by the Duke Endocarditis Service (Duke University, Durham, North Carolina, USA): positive blood cultures and echocardiogram (primary criteria); and fever, history of heart disease, recent heart murmur, and vascular and immunological phenomena (secondary criteria). A specific protocol was filled out by interns and medical students and revised by physicians from the hospital medical team to obtain the following data: age, sex, clinical and laboratory findings, and results of transthoracic echocardiography. In all cases, three blood samples were collected under aseptic conditions and inoculated in aerobic and anaerobic environments. The samples were then incubated at 37 degrees C and tested with the VITEC SYSTEM automatized system (BIOLAB). RESULTS: Among 28 patients, 16 were boys. Age was between 3 and 180 months (mean 70.6 +/- 59.2 months). Most children (68%) came from the metropolitan area of Vitória, and half presented a history of cardiopathy. The most frequent clinical findings which led to suspicion of IE were: prolonged fever in all cases, new or changing regurgitant murmur (67.9%), dyspnea (57.1%), hepatomegaly (57.1%), vascular phenomena (32.2%), splenomegaly (28.6%), and Osler's nodes (7.1%). Blood cultures were positive in 16 patients (57.1%), and community-acquired S. aureus was the most frequently isolated pathogen (56.6%). Despite the treatment, fever persisted for 2.5 to 30 days (median=18.0 days). Mean leukocyte count at admission was 11,657 +/- 7,085 mm3. Transthoracic echocardiography showed endocardial lesion in all patients, evidencing vegetations in the tricuspid valve (25%), mitral valve (25%), and intraventricular communication border (28%). Thirteen (46.4%) children presented concomitant sepsis and three (10.7%) developed hospital-acquired infection. There was one death (3.6%). CONCLUSIONS: IE in our country is frequent among children younger than 2 years with congenital cardiopathy. Community-acquired S. aureus was the most frequently isolated microorganism in children with no previous cardiac lesion. Acute infective endocarditis, bacterial endocarditis, Staphylococcus aureus, transthoracic echocardiography.

Adolescent↗
PubMed2004

The expression of CD44 in archival paraffin embedded interface tissues of failed orthopaedic implants.

CD44 hyaluronan receptor is present on large number of different cell type. It acts as one of the adhesion proteins, binding to hyaluronan and is known to play a part in cell migration from vessels in inflammation. The aim of this study was to examine the presence and distribution of CD44 in interface membrane in aseptic loosening. Immunohistochemistry (IHC) using human anti-mouse CD44 antibody studied 20 aseptically loosened interface samples. Extracted protein from all cases was examined by Western blot and RT-PCR. CD44 was detected in 85% of interfaces by IHC and the presence of protein confirmed by blotting and RT-PCR, which showed the mRNA level for CD44. CD44 was expressed by macrophage, multinucleated giant cells, mast cells and lymphocytes. Further studies are needed to characterise the role of this molecule in the inflammatory response to wear debris in aseptic loosening.

Aged↗
PubMed2005

Preventing bacterial contamination and proliferation during the harvest, storage, and feeding of fresh bovine colostrum.

The objectives of this study were to identify control points for bacterial contamination of bovine colostrum during the harvesting and feeding processes, and to describe the effects of refrigeration and use of potassium sorbate preservative on bacteria counts in stored fresh colostrum. For objective 1, first-milking colostrum samples were collected aseptically directly from the mammary glands of 39 cows, from the milking bucket, and from the esophageal feeder tube. For objective 2, 15-mL aliquots of colostrum were collected from the milking bucket and allocated to 1 of 4 treatment groups: 1) refrigeration, 2) ambient temperature, 3) refrigeration with potassium sorbate preservative, and 4) ambient temperature with potassium sorbate preservative. Subsamples from each treatment group were collected after 24, 48, and 96 h of storage. All samples underwent bacteriological culture for total plate count and coliform count. Bacteria counts were generally low or zero in colostrum collected directly from the gland [mean (SD) log10 cfu/mL(udder) = 1.44 (1.45)]. However, significant bacterial contamination occurred during the harvest process [mean (SD) log10 cfu/mL(bucket) = 4.99 (1.95)]. No additional bacterial contamination occurred between the bucket and the esophageal feeder tube. Storing colostrum at warm ambient temperatures resulted in the most rapid increase in bacteria counts, followed by intermediate rates of growth in nonpreserved refrigerated samples or preserved samples stored at ambient temperature. The most effective treatment studied was the use of potassium sorbate preservative in refrigerated samples, for which total plate count and total coliform counts dropped significantly and then remained constant during the 96-h storage period.

Animals↗
PubMed2003

Epidemiologic study of intramammary infections with Staphylococcus aureus during a control program in nine commercial dairy herds.

OBJECTIVE: To determine the epidemiologic pattern of intramammary infections (IMIs) with Staphylococcus aureus during implementation of a control program in 9 commercial dairy herds. DESIGN: Cohort study. ANIMALS: 1,651 lactating cows and 53,098 quarter milk samples. PROCEDURES: Nine herds located in different regions of Italy were enrolled. Control of S. aureus infections followed the general principles of contagious mastitis control and was based on precise diagnostic procedures and strict control and segregation of infected cows. All lactating cows in each herd were tested, and those free of S. aureus IMI were enrolled as the cohorts. Further additions to the cohort group were cows and heifers free of S. aureus IMI, as determined from aseptically collected milk samples taken approximately 7 and 14 days after calving. RESULTS: After the ninth month of the program, incidence decreased to < 2 new IMIs/100 cow-months in 7 of the herds. At the end of the study, 8 of 9 herds had an incidence of < or = 1 new IMI/100 cow-months. Heifers were most at risk of developing an IMI. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that control of S. aureus IMIs can be achieved by use of a control program based on segregation and use of hygienic and therapeutic protocols. Analysis of incidence and identification of risk factors in a herd could avoid the possible shortcomings of the program, maximizing the probability of success.

Animals↗
PubMed1996

The prevalence of mastitis in primiparous heifers in eleven Waikato dairy herds.

The objective of the study was to determine the prevalence of mastitis among primiparous heifers at calving and at drying off in 11 Waikato dairy herds during the 1993-94 dairy production season. Duplicate quarter milk samples were collected aseptically from 458 heifers within 5 days after calving for bacteriological analysis. Mastitis was diagnosed in at least one quarter in 35.6% of these heifers. Coagulase-negative staphylococci were isolated from 21.8% of the heifers. The prevalence of coagulase-negative staphylococci varied between herds from 4.3% to 44.8%. Environmental streptococci caused mastitis in 12.2% of heifers, ranging from 5.6% to 24.1% between herds. Streptococcus uberus was the pathogen identified most frequently at calving and accounted for more than 90% of the streptococcal isolates. Staphylococcus aureus and coliforms were isolated from less than 1% of samples. Clinical mastitis was observed in 8.1% of heifers at calving; environmental streptococci were isolated from 67.6% of these clinical clinical cases. Only 2.8% of heifers developed clinical mastitis during lactation and environmental streptococci were isolated from 38.5% of these cases. The prevalence of mastitis among 428 of the heifers at drying off was 64.7%; a 1.8 fold increase during lactation. Corynebactetium bovis was isolated from 43% of heifers at drying off even though it was not isolated from any heifers at calving. During the season, the prevalence of Staphylococcus aureus mastitis increased to 2.8% while mastitis caused by environmental streptococci declined to 2.8%. The prevalence of environmental mastitis pathogens decreased during lactation while contagious pathogens increased in each of the 11 herds. Ineffective post-milking teat sanitation probably contributed to the increase in mastitis caused by contagious pathogens. Specific factors were not determined that affected the variation in prevalence between herds.

Journal Article↗
PubMed1989

Keratinophilic fungi isolated from Antarctic soil.

In the present study, 10 soil samples were collected aseptically from an equal number of areas of the Antarctic in the zone occupied by the 1986-1987 Italian expedition for research on keratinophilic fungi. Of particular interest was the isolation of a pathogenic fungus, Microsporum gypseum, from two sites in the base camp occupied by men and by skuas. Trichophyton terrestre was isolated from a site in which people worked and through which penguins and skuas passed. The most widespread fungal species were members of the genus Chrysosporium. Some of these species were isolated but not identified and this part of the study was still be completed. Another significant finding was the absence of fungi in one sample, while in another the widespread and abundant growth in all the seeded dishes of a single species of Chrysosporium. Other species in genera of general diffusion in many environments were also isolated: Aspergillus spp., Malbranchea sp., Mycelia sterilia spp., Paecilomyces sp., Penicillium spp. and Scopulariopsis spp.

Animals↗
PubMed1993

Control of microbial activity by flow injection analysis during high cell density cultivation of Escherichia coli.

The application of an automated flow injection analysis (FIA) system for on-line determination of microbial activity, during high cell density cultivations of Escherichia coli is reported. Based on a bioelectrochemical principle, the FIA method used a redox mediator (potassium hexacyanoferrate(III)) to facilitate electron transfer from the microorganisms to an electrochemical detector. Assays were carried out using a new sampling device which provided aseptic operation by use of a valve and chemical sterilisation. No sample dilution or pretreatment was necessary for biomass concentrations up to approx. 40 g l-1. The sample volume was 0.5 ml and the overall analysis time was 5 min. FIA signals were found to correlate well with the oxygen uptake rate (OUR). Changes in metabolic activity due to low substrate levels or high inhibitor concentrations in the cultivation medium became obvious from the FIA signals.

Bacteriological Techniques↗
PubMed2004

Survival and injury of Arcobacter after artificial inoculation into drinking water.

The aim of this work was to assess the effect of chlorine water treatment on Arcobacter butzleri and to study the survival strategies of this organism in chlorinated and non-chlorinated drinking water. A. butzleri NCTC 12481 was inoculated into chlorinated and non-chlorinated water and samples were removed aseptically, immediately and periodically during the next 2 days (for chlorinated drinking water) or 35 days (for non-chlorinated drinking water). The membrane integrity (Live/Dead BacLight kit), 16S rRNA (FISH technique), DNA content (23S rRNA PCR-RFLPs) and culturability changes in A. butzleri cells were analyzed. Culturability of the cells was lost at 5 min in chlorinated drinking water. At that time the cells showed membrane damage, although fluorescent intensity of 16S rRNA hybridization was constant throughout the chlorine treatment. After 48 h the amplicon specific for the 23S rRNA gene was weakly detected. In non-chlorinated drinking water cells lost their culturability after 16 days but the other factors measured indicated that Arcobacter remained viable throughout the experiment.

Arcobacter↗
PubMed1999

Bacteremia after dental treatment in mentally handicapped people.

Bacteremia may occur after disruption of the oral mucous membrane, particularly after dental treatment. 18 mentally handicapped patients who underwent dental treatment with general anesthesia were included in our study. None of the patients had general illnesses or received antibiotic protection. From each patient several blood samples were drawn aseptically during dental treatment and cultured. The majority of aerobic bacteria recovered belonged to Streptococcus sp and Gemella sp., anaerobic bacteria mainly belonged to Porphyromonas gingivalis and Peptostreptococcus sp. Resistance of the isolated bacteria to penicillin as well as to oxacillin, erythromycin and Co-trimoxazole was substantial. The highest resistance rate could be shown against fucidic acid.

Adolescent↗
PubMed2000

Detection of bacterial virulence genes associated with infective endocarditis in infected root canals.

AIM: The aim of this study was to examine whether bacteria associated with root canals possess genes that might predispose to bacterial colonization of the endocardium. METHODOLOGY: Oligonucleotides were designed from DNA sequences encoding the functional binding regions of streptococcal fibronectin-binding protein (FnBP) and staphylococcal fibrinogen-binding protein (FgBP). The specificity and cross-reactivity of the oligonucleotide primers were investigated; streptococcal primers were tested for recognition of FnBP genes in other strains of streptococci, and the staphylococcal primers for detection of FgBP from other staphylococci. Interspecies specificity of these primers was also investigated. In a pilot clinical study, the pulp chambers of 16 nonvital teeth without sinus tracts, were opened aseptically. Root canal samples were collected, along with samples from the gingival sulcus and anterior nares. From these samples DNA was extracted, subjected to polymerase chain reaction (PCR) and analysed by agarose gel electrophoresis. RESULTS: Using the streptococcal FnBP primers, PCR bands were amplified from eight root canal samples, eight gingival samples and three nasal samples. With the staphylococcal primers, PCR bands were amplified from seven root canals, 11 gingival and nine nasal samples. This study showed that PCR could be used to detect bacteria in root canals that possess genes with homology to functional regions of those encoding FnBP or FgBP. CONCLUSIONS: If bacteria in root canals possess FnBP or FgBP, they may have the potential to cause infective endocarditis.

Adhesins, Bacterial↗
PubMed1992

Effects of prestorage white cell-reduction of apheresis platelets on platelet glycoprotein Ib and von Willebrand factor.

Twenty plateletpheresis components were harvested from 11 healthy donors and stored in polyolefin bags on a horizontal flatbed agitator at 22 degrees C. After 24 hours, white cells were reduced in one aliquot by centrifugation while the other aliquot was stored unaltered. Samples were obtained aseptically from each of these platelets at intervals for up to 10 days, and measurements were made of platelet glycoprotein Ib (GPIb) by both flow cytometry and polyacrylamide gel electrophoresis, of ristocetin-induced platelet aggregation by impedance aggregometry, and of plasma and platelet von Willebrand factor (vWF) by enzyme-linked immunosorbent assay. Storage of platelets under these conditions was associated with only minor decreases in surface GPIb, intraplatelet vWF, and ristocetin-induced platelet aggregation, and no differences were observed between the white cell-reduced and nonreduced aliquots. No benefit of white cell reduction in such components before prolonged storage is evident in the vWF-platelet interaction.

Blood Component Removal↗
PubMed1987

Management of infectious complications of intraventricular reservoirs in cancer patients: low incidence and successful treatment without reservoir removal.

At the time of analysis, the first 30 patients with Ommaya reservoirs (OR) at the Children's Orthopedic Hospital and Medical Center in Seattle, Washington had had 32 reservoirs for a mean duration of 28 months. In all, the reservoir chambers had been punctured for either diagnostic or therapeutic purposes a total of 1,287 times with a mean of 40 injections per reservoir and 1.4 injections per month. Six reservoir infections were diagnosed in five patients--a rate of one infection for every 153 reservoir-months. Four infections were attributed to reservoir use--a rate of one infection for every 322 reservoir entries, and less than one infection in 900 entries when a standard aseptic protocol for sampling and injection was applied. There was no correlation between infectious complications and the frequency with which reservoirs were injected, but there was evidence that some of the infections resulted from incomplete compliance with recommended technique for skin preparation and reservoir entry. Four infections were treated successfully with intravenous and intra-reservoir antibiotics without reservoir removal. Only one reservoir had to be removed because of persistent infection. Two other reservoirs were removed because of trauma and malfunction. Twenty-seven patients (90%) retained their original reservoir, up to 9.5 years after implantation. Intraventricular chemotherapy via an indwelling subcutaneous reservoir is a preferred method for delivery of intrathecal chemotherapy, with an acceptable infection risk relative to the benefits of patient comfort and therapeutic efficacy.

Adolescent↗
PubMed2001

Serum TGF-beta in thermally injured rats.

A severe thermal injury is commonly associated with immune suppression and increased susceptibility to sepsis, frequently leading to multiple organ failure. Transforming growth factor-beta (TGF-beta) is a potent immunosuppressive cytokine involved in complications associated with major trauma. Interleukin- 4 (IL-4) is thought to synergize the immunosuppressive activity of TGF-beta by promoting naive lymphocytes to differentiate and generate TGF-beta secreting cells. This study examines the alterations in serum levels of TGF-beta and IL-4 after a thermal injury. Male Sprague-Dawley rats (300-400 g) were anesthetized and received a 50% total body surface area full-thickness scald burn followed by fluid resuscitation and analgesia. Control rats were given the same treatment, but were immersed in water at room temperature. Rats were sacrificed from 1 h to 8 days after injury. Blood samples were collected aseptically from the inferior caval vein. Serum levels of TGF-beta and IL-4 were measured by enzyme linked immunosorbent assay. Rats in the control and thermal injury groups showed similar increases in serum TGF-beta 1 h after injury. A progressive increase in serum TGF-beta was observed in burned animals compared to control animals starting on day 3 and continued through day 8 (P < 0.01). Serum IL-4 levels in control and thermally injured animals remained undetectable (< 15.6 pg/mL) throughout the experiment. Thermal injury induces a significant increase in serum TGF-beta, which may contribute to post-burn immunosuppression with an increased susceptibility to sepsis.

Animals↗
PubMed1996

Anaerobic microorganisms in root canals of human teeth with chronic apical periodontitis detected by indirect immunofluorescence.

Aiming to assess the presence of selected anaerobic microorganisms in root canals of human teeth with chronic apical periodontitis. 25 central and lateral upper incisors presenting with radiographic evidence of chronic apical periodontitis were studied. The pulp chamber was opened under aseptic conditions and samples of the root canal content were collected with sterile absorbent paper points, which were placed and dispersed in test tubes containing reduced transport medium RTT. Aliquots were dried on glass slides and stained by indirect immunofluorescence for detection of Actinomyces viscosus, Fusobacterium nucleatum, Porphyromonas gingivalis and Prevotella intermedia. The results showed a positive indirect immunofluorescence reaction in 24 of the 25 samples. Fourteen were positive for the specie Actinomyces viscosus, 12 for Prevotella intermedia, 10 for Fusobacterium nucleatum and 4 for Porphyromonas gingivalis. A semiquantitative assay was easily implemented for assessment of degree of infection by the organisms in individual cases.

Actinomyces viscosus↗
PubMed1971

Incidence of prosthecate bacteria in a polluted stream.

Water samples were collected aseptically several times throughout the year at nine stations on the Red Cedar River, a stream flowing through farmland and receiving effluent from several municipalities in central Michigan. Total prosthecate bacteria were enumerated by both direct and viable counting techniques. By direct techniques, these bacteria accounted for 0.62 to 1.1% of the total microflora during the study. The predominant type of appendaged bacteria was the caulobacters (Caulobacter, Asticcacaulis, and the fusiform caulobacter), which accounted for 64 to 93% of the total prosthecate forms. The others of importance were prosthecomicrobia (< 1 to 24%), including Prosthecomicrobium and Prosthecochloris; hyphomicrobia (< 1 to 15%), including Hyphomicrobium and Rhodomicrobium; and Ancalomicrobium (< 1 to 6%). The viable counts of heterotrophs also indicated that the caulobacters were the most numerous prosthecate bacteria in the stream. They ranged from fewer than 1 per ml to a maximum of almost 4,000 per ml. During the coldest period, when the total viable counts decreased to about 10(4) per ml compared to their summer high of over 10(7) per ml, the caulobacters actually increased in numbers. In December (temperature 0 to 1 C), they comprised from 0.09 to 1.0% of the viable microbial count, and in March (6.0 to 8.0 C) they accounted for 0.14 to 2.8%. The other heterotrophic prosthecate bacteria were generally found at numbers less than 1 per ml, with the exception of the December study when Hyphomicrobium was present in numbers as high as 2,400 per ml. There was no consistent correlation between the frequency of prosthecate bacteria and total coliforms in the stream during the investigation.

Bacteria↗
PubMed2001

Effects of amphotericin B gargles on oral colonization of Candida albicans in asthmatic patients on steroid inhalation therapy.

BACKGROUND: Early use of inhaled steroids is recommended for bronchial asthma. The side effects are rare, but oral discomfort and candidiasis are clinically important complications. Most previous studies reported that the use of spacer and water gargling was necessary to prevent oral complications. However, in some patients, this may fail to prevent such complications. OBJECTIVE: To compare the effects of water gargling with those of amphotericin B, in the prevention of oral complications in asthmatics using inhaled steroids. METHODS: Pharyngeal swab samples were obtained aseptically from the posterior pharyngeal wall of 128 asthmatics who have been using inhaled steroids (beclomethasone dipropionate) for more than 1 year. The amount of Candida albicans in cultured swabs was evaluated based on the following criteria: oral symptoms, method of gargling, dose of inhaled steroids, type of spacer and serum cortisol level. RESULTS: The number of isolated C. albicans was significantly higher in asthmatics with oral symptoms than in those free of symptoms. It was also significantly higher in patients who gargled with water or 1,000 times dilution than in those who gargled with 100 or 50 times dilutions of amphotericin B. Moreover, it was significantly higher in patients with low levels of serum cortisol than in those with normal serum cortisol. CONCLUSION: We demonstrated that at least in a subgroup of asthmatics using steroid inhalers, gargling with water or even weak concentrations of amphotericin B does not prevent colonization of the throat with C. albicans. This group at high risk of developing oral candidiasis should gargle with amphotericin B at concentrations higher than 100 times dilution that can prevent clinically detectable oral candidiasis.

Administration, Oral↗
PubMed2003

Isolation of an unusual fungus in treated dental unit waterlines.

BACKGROUND: Numerous organisms have been identified in dental unit waterlines, or DUWLs. Decontamination of DUWLs focuses on maintaining heterotrophic, mesophilic bacteria below 200 colony-forming units per milliliter as recommended by the ADA. METHODS: The authors conducted a study to test the efficacy of a continuous-use, stabilized chlorine dioxide proprietary compound to decrease the number of bacteria in DUWLs. The authors used three dental units with self-contained water systems to test the product and three similar units as controls. They aseptically collected water samples weekly according to recommended methods, plated the samples on R2A agar and incubated them for seven days. RESULTS: The authors isolated heterotrophic, mesophilic bacteria from treatment and control units for eight weeks. In the ninth week, the predominant isolates from one of the treatment units changed in appearance to small, dark, shiny colonies that the authors tentatively identified as fungal. The authors then isolated similar colonies from the source tap water and ultrasonic and handpiece lines. They added three additional dental units from the same clinic in the sixth week of the study and isolated similar fungal colonies from them after five weeks of treatment. The authors performed DNA sequencing with an automated sequencer and identified the organism Exophiala mesophila. CONCLUSIONS: The authors did not observe fungal isolates in the control units, which suggests that continuous waterline treatment may cause proliferation of a fungus present in small amounts in source water. CLINICAL IMPLICATIONS. The findings of this study indicate the need to monitor water quality regularly when treating waterlines with continuous-use chemical cleaners.

Bacteria↗
PubMed1976

E. coli growth inhibition by amniotic fluid.

Amniotic fluid samples were collected aseptically from 29 normal pregnancies, between the 38th and 41st week of gestation and checked for sterility in the laboratory, in order to investigate in vitro the effect of the liquor on the growth of E. coli. Brain Heart Infusion (B.H.I.) and Ringer solution were used as controls. E. coli cultures were inoculated in amniotic fluid, B.H.I., Ringer solution and amniotic fluid plus B.H.I. and incubated at 37degreesC for 48 hours. At 0, 1, 3, 6, 12, 24, 36 and 48 hours of incubation, surface viable counts were performed to estimate the number of E. coli viable cells. The growth of E. coli in B.H.I. started furing the 1st hours after inoculation and continued over 48 hours; in Ringer solution the mean growth curve and almost identical to that of B.H.I. In amniotic fluid the growth of E. coli began from the 1st hour of inoculation but the growth curve was much lower, became static in 24 hours and a permanent inhibition was observed thereafter. The addition of a small amount of B.H.I. in amniotic fluid enhanced the growth of E. coli, but the growth curve was lower in comparison to the curves of the two controls studied. In conclusion, after 48 hours of inoculation in amniotic fluid, 26 of the 29 cases showed bacteriostatic or bactericidal activity of the liquor upon the growth of E. coli.

Amniotic Fluid↗