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Bacterial safety of a closed-administration system for enteral nutrition solutions.

The purpose of this study was to investigate the bacterial integrity of a newly designed closed-administration system for enteral formula delivery. Three clinical simulations, designated Phases I, II, and III, were tested: dispersal of (I) 3 L formula over a 24-hour period, (II) 2 L formula over a 24-hour period, and (III) 2 L formula over a 48-hour period. Within each 24- or 48-hour simulation phase, a single administration set was used. Samples were withdrawn for bacterial analysis at 4- or 8-hour intervals. Simulations and samplings were completed under controlled, but not aseptic, conditions. Results indicated that bacterial growth was insignificant across all three simulation phases. Re-utilization of the administration set did not introduce bacterial contamination into subsequent liters of formula. Alcohol cleansing of the administration set was not found to be necessary for maintenance of bacterial closed integrity. Even when administered over a 24- or 48-hour period, the formula dispersed through this closed system did not develop any significant degree of bacterial growth. Results of this study suggest that this newly designed system for the administration of enteral formula significantly minimizes the risk of bacterial contamination.

Bacteria↗

Mechanical characteristics of bone in femoral capital aseptic necrosis.

Uniaxial compression loading of small cube-shaped samples in an Instron machine was used to study material property deficits of middle- and late-stage necrotic cancellous bone taken from adult human femoral heads. Although the experimental data should not be extrapolated to phenomena of viable trabecular response in a functioning osteonecrotic hip, they provide insight into the salient passive mechanical response characteristics of the pathologic tissue. Individual differences in the revascularization and/or collapse patterns occurring within femoral heads lead to substantial variability in the strength and stiffness of test samples taken from the involved regions. As an overall average, however, samples extracted from major infarct regions exhibit substantially reduced (52%) yield strength, severely reduced (72%) elastic modulus, and modestly increased (29%) strain-to-failure, relative to samples from normal femoral heads. However, statistically significant changes are not observed in the Poisson ratio, the post-yield modulus, or in the incidence of pre-yield load/deformation discontinuities.

Biomechanical Phenomena↗

[The protective effects and its underlying mechanism of 2,4-diamino-6-hydroxy-pyrimidine on postburn Staphylococcus aureus sepsis in rats].

OBJECTIVE: To investigate the protective effect and its underlying mechanism of 2,4-diamino-6-hydroxy-pyrimidine (DAHP), an inhibitor of GTP-cyclohydrolase I (GTP-CHI), on postburn Staphylococcus aureus (S. aureus) sepsis in rats. METHODS: Fifty-six Wistar rats were randomly divided into four groups, i.e. normal control, scalding control, postburn sepsis group and DAHP treatment group. Tissue samples from liver, kidneys, lungs and heart were aseptically taken, and in which the GTP-CHI and inducible nitric oxide synthase (iNOS) contents and the mRNA expression of tumor necrosis factor-alpha (TNFalpha) were determined. Furthermore, biopterin (BH(4)) and nitric oxide (NO) levels in these tissue were also measured. RESULTS: After the scalding injury followed by bacterial challenge, the GTP-CHI gene expression and biopterin levels were significantly increased in all tissue sampled, and so were iNOS mRNA expression and NO (P < 0.01), especially in liver and lungs. The expressions of GTP-CHI mRNA and iNOS mRNA and the production of BH(4) and NO in all tissue were evidently inhibited by the pretreatment with DAHP (P < 0.05 approximately 0.01). At the same time, the TNFalpha expression was also obviously decreased. In addition, The mortality at 6 hr in rats of DAHP treatment group was decreased. CONCLUSION: The prognosis of the scalding rats complicated by sepsis caused by G(+) bacteria could be improved by DAHP pretreatment, which might be related to the inhibition of the production of BH(4) and NO by DAHP.

Animals↗

Microbiological consequences of skin removal prior to evisceration of broiler carcasses.

The objective of this project was to determine if removal of skin prior to evisceration lowers the number of bacteria that can be recovered by whole carcass rinse or sponge sampling. Four experiments were conducted, two with each type of sampling (rinse or sponge). New York dressed carcasses obtained from a commercial broiler processing plant were aseptically skinned or left with skin intact. The carcasses were then aseptically eviscerated by hand. Carcasses were rinsed in 100 mL sterile water or sampled by moist sponge. When sampled by rinse, significantly fewer Campylobacter and total aerobic bacteria were recovered from carcasses that had been skinned prior to evisceration. When sampled by sponge, significantly fewer Campylobacter, Escherichia coli, coliform and total aerobic bacteria were recovered from the outer surface of carcasses without skin. No differences were noted for bacterial counts recovered from internal surfaces by sponge sampling. Similar trends were observed when carcasses were subjected to an inside and outside washing step after evisceration. Removal of skin and washing the carcass led to significantly less Campylobacter being recovered by whole carcass rinse compared to carcasses that were washed with the skin on. When sampled by sponge, incidence of Campylobacter and level of total aerobic bacterial counts were lower on the outer surface of skinned and washed carcasses than on washed carcasses with intact skin. Like the unwashed carcasses, no differences were noted for bacterial counts recovered from internal surfaces by sponge sampling. Although not commercially practical, it is possible to lower the level of Campylobacter on the outside of broiler carcasses by removal of the skin prior to evisceration.

Animals↗

Persistent increase of matrix metalloproteinases in cerebrospinal fluid of tuberculous meningitis.

Matrix metalloproteinase (MMP)-2 and MMP-9 were analyzed by gelatin zymography and an enzyme-linked immunosorbent assay (ELISA) in a cerebrospinal fluid (CSF) from patients with tuberculous meningitis (n=24), acute aseptic meningitis (n=23) and the control (n=10). The MMP-2 and MMP-9 levels were significantly higher in the samples from the tuberculous meningitis patients than those from either the aseptic meningitis patients or the controls. In tuberculous meningitis, the patients with late neurologic complications had higher MMP-2 and MMP-9 levels than those without. The persistent increase in the MMP-2 and MMP-9 levels was associated with the development of complications following tuberculous meningitis. Inhibiting the MMPs may be an effective strategy for preventing or reducing the complications in tuberculous meningitis.

Electrophoresis, Polyacrylamide Gel↗

Cerebrospinal fluid leukocyte aggregation in meningitis.

OBJECTIVE: To evaluate whether the difference in aggregation of cerebrospinal fluid cells from patients with bacterial, viral, aseptic and partially treated meningitis can be used for diagnostic purposes. METHODS: Cerebrospinal fluid samples of 100 patients with meningitis (15 bacterial, 13 partially treated, 10 viral and 62 aseptic) were compared on the basis of the predefined leukocyte aggregation score (LAS). RESULTS: Mean LAS was 56% in the bacterial meningitis group (range, 15 to 90%), 5.8% in the partially treated meningitis group (range, 0 to 27%), 2% in the proven viral meningitis group (range, 0 to 5%) and 2% in the aseptic meningitis group (range, 0 to 15%). All patients with bacterial meningitis had a LAS of > 15%, whereas all those with viral or aseptic meningitis had a score of < 15%. Although most patients with partially treated meningitis had a low LAS, several had higher scores, which may indicate bacterial infection. There was no statistical correlation between number of cells, type of cells (mononuclear or polymorphonuclear) or cerebrospinal fluid protein and glucose concentration and degree of leukocyte aggregation for the different groups. CONCLUSION: Measurement of the LAS may contribute to the immediate differential diagnosis of bacterial or viral meningitis, especially in patients with very high pleocytosis, as sometimes seen in enteroviral meningitis. It may also serve as a guide for the likelihood of bacterial infection in cases of partially treated meningitis. Additional studies are needed to confirm these observations.

Adolescent↗

CSF lymphocyte subsets in aseptic meningitis: dual-labelling analysis with flow cytometry.

In order to characterize the CSF (cerebrospinal fluid) lymphocytes in CNS (central nervous system) inflammation, we examined paired samples of CSF and PB (peripheral blood) of 19 patients with acute aseptic meningitis, performing the dual labelling method on flow cytometry. Significantly higher percentages of CD3+ (T cell), CD4+ (helper-inducer), Leu3a+ Leu15- (cytotoxic-T) and HLA-DR+ CD3+ (activated-T) cells were identified in the CSF than in the PB of these patients. We observed significantly lower percentages of CD19+ (B cell), Leu2a+ Leu18+ (suppressor-inducer) and HLA-DR+ CD3- cells in the CSF than in the PB of these patients. Relative increases in helper-inducer, cytotoxic-T and activated-T cells in the CSF of aseptic meningitis are supposed to represent an active inflammatory process. However, whether these changes are specific or pathognomonic to any disease(s) remains to be solved.

Adolescent↗

Focal dermatitis and cellulitis in broiler chickens: bacteriological and pathological findings.

Involvement of aerobic bacteria, especially Escherichia coli, in lesions of cellulitis in broiler chickens was investigated. Samples of subcutaneous caseous material for bacteriological examination were collected aseptically at the slaughterhouse from 109 broilers with lesions. Skin tissue was taken from five of these birds for histopathological examination. In 96 of the 109 (88.1%) broilers sampled, E. coli was isolated from the lesion, and in 60 of these birds it was the only bacterial species found. In 33 broilers, Streptococcus dysgalactiae was found along with E. coli. Although serotype O78 was isolated the most frequently, numerous other serotypes were found; no correlation could be established between the size of the lesions and the serotype isolated. The microscopic lesions were characterized by thickening of the dermis with a granulomatous inflammatory reaction. This study confirms the frequent association of E. coli with cellulitis lesions in broiler chickens and reports the frequent isolation of S. dysgalactiae from the lesion.

Animals↗

[Study of clean versus aseptic technique of tracheotomy care based on the level of pulmonary infection].

The purpose of this research study was to determine whether the clean technique of tracheotomy care is the same, or more, secure from the aseptic, by testing the difference in the level of postoperative pulmonary infection between tracheotomized patients receiving aseptic and those receiving clean tracheotomy care. The sample consisted of 103 patients with tracheotomy (transient or permanent) from ENT or IC units of four big hospitals of Athens. The level of the patient's pulmonary infection was defined using the Weighted Level of Pulmonary Infection Tool, which was constructed especially for this research study. The data were analysed using the x2 statistical test, and the coefficients phi, Cramer's V and Kendall's, while, with the same statistics, the relationship between certain important external variables and the dependent variable was examined. The findings indicated that no statistically significant difference exists between clean and aseptic technique as to the level of pulmonary infection when used for tracheotomy postoperative care of tracheotomized patients.

Adult↗

Subclass distribution of the serum and intrathecal IgG antibody response in varicella-zoster virus infections.

The subclass distribution of the varicella-zoster virus (VZV)-specific IgG antibody response was studied in serum samples from 22 patients with primary varicella and 34 with recurrent VZV infections and in cerebrospinal fluid (CSF) samples from 22 patients with recurrent infection who presented with symptoms of aseptic meningitis. IgG1 and 3 were the dominant subclasses among patients with primary and recurrent infections; IgG1 was also prevalent in the CSF samples. The VZV IgG subclass distribution patterns did not allow differentiation between primary and recurrent infections. However, seroconversions for IgG2, 3, or 4 were observed among patients with recurrences who were negative for specific IgM, suggesting that qualitative tests for serum IgG subclass antibody could be helpful for diagnosis in such cases. Herpes simplex virus-specific IgG was found in CSF samples from several patients with meningitis. The results suggest that calculation of the antibody to albumin index is better than IgG subclass antibody assays for discriminating the causative agent in these cases.

Antibodies, Viral↗

Production of platelet-derived growth factor in aseptic loosening of total hip replacement.

Aseptic loosening is the predominant cause of total hip implant failure. It has been assumed that a layer or membrane, containing macrophages, fibroblasts and vascular endothelial cells, of synovial-like tissue develops at the implant-to-bone interface almost invariably and, with time, somehow leads to loosening of the components from the surrounding bone. These cells produce a variety of cytokines and proteolytic enzymes which stimulate bone resorption. Platelet derived growth factor (PDGF) may be one of the cytokines which stimulate bone resorption and contribute to aseptic loosening in total hip replacement (THR). Synovial-like membrane from the implant or cement-to-bone interface (n = 10) and pseudocapsule (n = 10) were obtained from ten patients operated on for aseptic loosening of THR. As a control, nine samples of connective tissues were obtained from patients who had mandibular or maxillary fractures fixed with bone implant. The avidin-biotin-peroxidase complex (ABC) method with polyclonal rabbit anti-human IgG against the A-chain and B-chain of PDGF was used for staining. ABC-alkaline phosphatase-anti-alkaline-phosphatase double staining with monoclonal mouse anti-human fibroblast IgG1 and CD68 antibodies was used to ascertain the cellular origin of PDGF. Results of the PDGF staining were quantitated by a semi-automatic VIDAS image analysis system. The PDGF-A and PDGF-B chain containing cells were found in all periprosthetic tissues, in particular in macrophages with phagocytosed particulate debris, but to some extent also in fibroblasts and in endothelial cells. The numbers of PDGF-A and PDGF-B chain positive cells per mm 2 in synovial-like interface membrane (1881 +/- 486 and 1877 +/- 214) and pseudocapsule (1786 +/- 236 and 1676 +/- 152) were higher (P < 0.01) around loose THR than in control tissue (821 +/- 112 and 467 +/- 150), respectively. The results of the present study suggest that PDGF is preferably expressed by macrophages, which to an increased extent produce it in the synovial-like interface membrane and pseudocapsular synovial-like membrane. Because of its role in bone resorption, it may well play a role in periprosthetic bone loss and aseptic loosening and deserves more detailed study as a mediator and potential target in the modulation or prevention of loosening of THR.

Adolescent↗

The diagnosis of tuberculous meningitis using the polymerase chain reaction.

AIM: DNA amplification by the polymerase chain reaction (PCR) was evaluated as a means for rapid diagnosis of tuberculous meningitis (TBM). METHODS: A 240 bp region (nts 460-700) from the MPB 64 protein coding gene specific for Mycobacterium tuberculosis (TB) was selected for amplification. Nineteen clinical samples were studied. Six were obtained from patients with TBM diagnosed by culture (4/6) or by response to therapy (2/6). The remaining 13 samples were obtained from patients with febrile seizu es (8/13), aseptic meningitis (3/13) and septic meningitis (2/13), and these served as negative controls. RESULTS: We detected TB DNA in all the 6 CSF specimens obtained from patients with TBM. PCR alone was sufficient to detect TB DNA in 5 of these 6 samples. However, one sample was positive only when PCR was followed by oligonucleotide hybridisation. In the 2 patients whose CSF were obtained only after commencement of TB therapy, TB cultures were negative but positive on PCR nd oligoprobe labelling. The diagnosis of TBM was confirmed based on their remarkable response to therapy. Twelve of the thirteen negative controls were TB DNA negative. There was one false positive sample, which was thought to be due to TB DNA contamination. CONCLUSION: Taken together, our results indicate that DNA amplification using PCR, followed by oligonucleotide hybridisation offers a rapid (5 working days) means of diagnosis of TBM, provided care is taken to ensure that cross contamination of DNA samples is avoided.

DNA Primers↗

Attached and unattached bacterial communities in a 120-meter corehole in an acidic, crystalline rock aquifer.

The bacteria colonizing geologic core sections (attached) were contrasted with those found suspended in the groundwater (unattached) by examining the microbiology of 16 depth-paired core and groundwater samples using a suite of culture-independent and culture-dependent analyses. One hundred twenty-two meters was continuously cored from a buried chalcopyrite ore hosted in a biotite-quartz-monzonite porphyry at the Mineral Park Mine near Kingman, Ariz. Every fourth 1.5-m core was acquired using microbiologically defensible methods, and these core sections were aseptically processed for characterization of the attached bacteria. Groundwater samples containing unattached bacteria were collected from the uncased corehole at depth intervals corresponding to the individual cores using an inflatable straddle packer sampler. The groundwater was acidic (pH 2.8 to 5.0), with low levels of dissolved oxygen and high concentrations of sulfate and metals, including ferrous iron. Total numbers of attached cells were less than 10(5) cells g of core material(-1) while unattached cells numbered about 10(5) cells ml of groundwater(-1). Attached and unattached acidophilic heterotrophs were observed throughout the depth profile. In contrast, acidophilic chemolithotrophs were not found attached to the rock but were commonly observed in the groundwater. Attached communities were composed of low numbers (<40 CFU g(-1)) of neutrophilic heterotrophs that exhibited a high degree of morphologic diversity, while unattached communities contained higher numbers (ca. 10(3) CFU ml(-1)) of neutrophilic heterotrophs of limited diversity. Sulfate-reducing bacteria were restricted to the deepest samples of both core and groundwater. 16S ribosomal DNA sequence analysis of attached, acidophilic isolates indicated that organisms closely related to heterotrophic, acidophilic mesophiles such as Acidiphilium organovorum and, surprisingly, to the moderately thermophilic Alicyclobacillus acidocaldarius were present. The results indicate that viable (but possibly inactive) microorganisms were present in the buried ore and that there was substantial distinction in biomass and physiological capabilities between attached and unattached populations.

Colony Count, Microbial↗

Mastitis in beef cows and its effects on calf weight gain.

Quarter milk samples from 51 purebred (Angus, Polled Hereford, and Simmental) and 69 crossbred (Angus x Simmental x Charolais three-way cross) beef cows were collected aseptically at three times during lactation to determine the prevalence of intramammary infection, milk somatic cell counts (SCC), and effects of infection on calf weight gain. Quarter infection prevalence was 13.1, 14.9, and 27.5% in early, mid, and late lactation; corresponding cow infection prevalence was 25.8, 29.2, and 54.4%. Staphylococcus aureus was isolated from 2.9, 2.7, and 3.2% of quarters in early, mid, and late lactation, respectively. Corynebacterium bovis, generally regarded as a minor pathogen, was isolated from 4.0, 7.6, and 18.2% of quarters at the three respective times. Geometric SCC means (10(3) cells/ml) were 1,522, 344, and 509 for S. aureus-infected quarters; 344, 899, and 221 for Staphylococcus hyicus-infected quarters; 65, 36, and 86 for C. bovis-infected quarters; and 20, 17, and 18 for uninfected quarters in early, mid, and late lactation, respectively. Adjusted 205-d weight gain for calves with S. aureus-infected dams was 9.6 kg less (P less than .05) than for calves with uninfected dams. Adjusted 205-d weight gain for calves with dams infected with any mastitis pathogen did not differ significantly from that of calves with uninfected dams. At weaning half of the infected cows and half of the uninfected cows were given an intramammary infusion product containing 300 mg of cephapirin benzathine in each quarter; the remaining cows were untreated controls. Quarter samples were collected aseptically from all cows 14 to 28 d after subsequent calving. Quarter prevalence of infection after calving was lower (P less than .05) in treated (8.2%) than in control (22.4%) cows. Significantly more infections present at weaning were eliminated in treated than in control cows, but the new infection rate during the dry period and early lactation did not differ between the two groups.

Age Factors↗

Prevalence of pigs infected by Salmonella Typhimurium at slaughter after an enterocolitis outbreak.

A cross-sectional study was performed to estimate the prevalence of slaughter pigs infected by Salmonella typhimurium after an enterocolitis outbreak in a commercial pig farm, which was characterised by diarrhoea during the growing phase. Anatomopathological and histopathological findings were suggestive of salmonellosis, which was further confirmed by isolation of S. typhimurium from organs and faeces samples from diseased animals. Ileocolic lymph nodes were aseptically collected from 43 pigs during slaughter procedures. The estimated prevalence of Salmonella-infected pigs was 53.48% [confidence interval (CI): 42.94:64.02%]. This finding demonstrates that the carriage of S. typhimurium at slaughter might be high if pigs originate from a batch previously affected by Salmonella-enterocolitis outbreak at the pre-harvest pork production chain.

Abattoirs↗

Nitric oxide synthase in tissues around failed hip prostheses.

Nineteen patients who had undergone hip revision surgery for aseptic loosening of joint prostheses were studied. Tissue samples were harvested at the interface between bone and implant, either at the stem or at the cotyle level. Immunohistochemistry was performed on tissue sections to detect nitric oxide synthase (NOS), the enzyme which enables the synthesis of nitric oxide (NO), a molecule which can activate bone resorption. Quantitative analysis of the positive cells and correlation with the presence of particulate wear debris and radiological data were performed. The authors observed a trend towards a moderate increase in positive cells due to inducible NOS in tissues containing particulate wear debris, especially of a plastic material. This increase, however, did not achieve statistical significance. On the contrary, there was a statistical correlation between iNOS (inducible NOS) and the severity of osteolysis around the prosthetic implant. Pharmacological control of the biosynthesis of NO may be considered in the prevention or treatment of loosening.

Adult↗

Inactivation of the bactericidal activity of human serum by liquoid (sodium polyanetholsulfonate).

Four serum-sensitive strains of Escherichia coli were exposed to 10, 20, and 50% fresh, heat-inactivated, and fresh human serum to which had been added Liquoid at a final concentration of 0.05, 0.025, 0.0125 and 0.006%. It was found that 50% fresh serum (in nutrient, Mueller-Hinton, thioglycolate, or Trypticase Soy Broth) killed more than 10(4) organisms/ml within 3 min, whereas 20 and 10% fresh serum required up to 20 and 40 min, respectively, to kill a comparable number of organisms. To neutralize the activity of 50% fresh serum, 0.0125% Liquoid had to be added, whereas an 0.006% final concentration of Liquoid was sufficient to antagonize the activity of 10 and 20% serum. However, when exposing extremely small bacterial inocula to fresh serum, at least 0.025% Liquoid was necessary to abolish the serum-bactericidal activity of 20 and 50% fresh serum. Liquoid had to be added to 50% fresh serum within seconds to prevent the killing of the majority of test organisms derived from small inocula. It is recommended that blood samples drawn from septicemic or bacteremic patients be aseptically added to a suitable broth which contains at least 0.025% Liquoid in order to improve the chances of isolating pathogens present in small numbers.

Bacteriological Techniques↗