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PubMed2005

Pathogen survival during livestock manure storage and following land application.

This paper reports the first year results of field experiments to determine the survival times of pathogens in livestock manures during storage and following land application, using viable count methods. E. coli O157, Salmonella and Campylobacter survived in stored slurries and dirty water for up to three months, with Listeria surviving for up to three months. In contrast, all these pathogens survived for less than one month in solid manure heaps where temperatures greater than 55 degrees C were obtained. Following manure spreading to land, E. coli O157, Salmonella and Campylobacter generally survived in the soil for up to one month after application to both the sandy arable and clay loam grassland soils, whereas Listeria commonly survived for more than one month. These data are being used to develop guidelines on the management of manures to minimize the risks of pathogen transfer from animal manures to the human food chain.

Agriculture↗

PubMed2001

Application of digital image analysis and flow cytometry to enumerate marine viruses stained with SYBR gold.

A novel nucleic acid stain, SYBR Gold, was used to stain marine viral particles in various types of samples. Viral particles stained with SYBR Gold yielded bright and stable fluorescent signals that could be detected by a cooled charge-coupled device camera or by flow cytometry. The fluorescent signal strength of SYBR Gold-stained viruses was about twice that of SYBR Green I-stained viruses. Digital images of SYBR Gold-stained viral particles were processed to enumerate the concentration of viral particles by using digital image analysis software. Estimates of viral concentration based on digitized images were 1.3 times higher than those based on direct counting by epifluorescence microscopy. Direct epifluorescence counts of SYBR Gold-stained viral particles were in turn about 1.34 times higher than those estimated by the transmission electron microscope method. Bacteriophage lysates stained with SYBR Gold formed a distinct population in flow cytometric signatures. Flow cytometric analysis revealed at least four viral subpopulations for a Lake Erie sample and two subpopulations for a Georgia coastal sample. Flow cytometry-based viral counts for various types of samples averaged 1.1 times higher than direct epifluorescence microscopic counts. The potential application of digital image analysis and flow cytometry for rapid and accurate measurement of viral abundance in aquatic environments is discussed.

Cyanobacteria↗

PubMed1977

[Contribution to the experimental haemophilus infection (haemophilus parahemolyticus, haemophilus parasuis) in specific pathogen-free piglets. 1. microbiology, experimental arrangement, results].

Experimental infections were applied to specific pathogen free (SPF) piglets and store pigs, using five haemophilus (H.) parahaemolyticus and two H.-parasuis strains. Different germs counts and modes of application were chosen for each of the animals involved (intratracheal, intranasal, and subcutaneous routes). Clinical and pathologico-anatomic changes typical of haemorrhagic-necrotising pleuropneumonia were obtained from all germ counts and methods of application. Only one of the test animals could be successfully infected to exhibit manifestations of pneumonia or serositis when H. parasuis was used. The high pathogenicity of H. parahaemolyticus, as recorded from the above experiments, was in agreement with the growing incidence of that haemophilosis recordable for some time from pig stock.

Animals↗

PubMed1989

Effectiveness of measures requiring and not requiring prior sequence alignment for estimating the dissimilarity of natural sequences.

Various measures of sequence dissimilarity have been evaluated by how well the additive least squares estimation of edges (branch lengths) of an unrooted evolutionary tree fit the observed pairwise dissimilarity measures and by how consistent the trees are for different data sets derived from the same set of sequences. This evaluation provided sensitive discrimination among dissimilarity measures and among possible trees. Dissimilarity measures not requiring prior sequence alignment did about as well as did the traditional mismatch counts requiring prior sequence alignment. Application of Jukes-Cantor correction to singlet mismatch counts worsened the results. Measures not requiring alignment had the advantage of being applicable to sequences too different to be critically alignable. Two different measures of pairwise dissimilarity not requiring alignment have been used: (1) multiplet distribution distance (MDD), the square of the Euclidean distance between vectors of the fractions of base signlets (or doublets, or triplets, or ...) in the respective sequences, and (2) complements of long words (CLW), the count of bases not occurring in significantly long common words. MDD was applicable to sequences more different than was CLW (noncoding), but the latter often gave better results where both measures were available (coding). MDD results were improved by using longer mutliplets and, if the sequences were coding, by using the larger amino acid and codon alphabets rather than the nucleotide alphabet. The additive least squares method could be used to provide a reasonable consensus of different trees for the same set of species (or related genes).

Animals↗

PubMed1999

Development of a surface adhesion immunofluorescent technique for the rapid detection of Salmonella spp. from meat and poultry.

A rapid method based on bacterial adhesion was developed for the detection of Salmonella in an enriched meat system. Minced beef samples inoculated with Salm. enteritidis (10 cfu g-1) were incubated overnight (18 h) at 37 degrees C in buffered peptone water. Salmonella enteritidis cells were isolated from the enriched meat sample by surface adhesion onto a polycarbonate membrane attached to a glass slide. The organisms attached to this polycarbonate membrane were subsequently visualized using immunofluorescent microscopy. The technique had a detection level of log10 3.5 Salmonella ml-1. The surface adhesion immunofluorescent technique correlated well with Salmonella plate counts (r2 = 0.99). Application of the rapid method to retail beef and poultry samples (n = 100) confirmed the correlation between this technique and traditional microbiological procedures. Thirty-one retail samples were reported positive for Salmonella species. No false positives or negatives were recorded for the rapid method.

Animals↗

PubMed1976

Counting losses in multidetector radiation counters.

We examined the reasons for the reduction in response of a multidetector radiation counting system when operated at high count-rates and established a convenient method of correction which is accurate to within +/- 1% up to 0-6 X 10(5) counts per second. In its application the method uses the observed count-rates from each detector at the time of measurement and previously determined values of critical time intervals of the electronic system. The detected signals establish these intervals in which further signals occurring within them are lost from the response. These further signals can also cause the loss of the signals establishing the intervals. The correction is of particular value when high-sensitivity whole-body counters are used to measure the activity of administered isotopes in metabolic studies.

Radiometry↗

PubMed1978

Computers and quality control in nuclear medicine.

The general topic of computers and nuclear medicine quality control may be approached from two main areas; controlling the quality of computerized studies, and computer applications in general nuclear medicine quality control. Overlap occurs when quality control of computer studies is performed by the computer itself. The uses of computers in record-keeping and in quality control of imaging instrumentation and in vitro studies, including radioimmunoassay, are discussed in this review. Aspects of quality control for computerized clinical cardiovascular, cerebral, and renal studies and emission computed tomography are reviewed, including consideration of difficulties and inaccuracies involved in the studies. Any automatic computer analysis program should incorporate adequate checks and error detection protocols and should illustrate results for verification. Current routine quality control procedures using the computer unfortunately are few. Quality control criteria are needed for camera/computer systems in high count rate clinical applications, and increasing emphasis should be aimed at quality control of those computerized dynamic and function studies in current clinical use. The computer has a valuable potential for nuclear medicine quality control. In vitro and computerized in vivo studies can be analyzed by readily available statistical programs, and variances can be monitored continuously. Computers can calibrate and monitor instrument performance regularly, and can handle managerial and clerical duties such as bookkeeping.

Cardiovascular Diseases↗

PubMed1989

Observations on the anaerobic membrane-filter technique for coliform counts in Nigerian surface waters.

The applicability of the anaerobic membrane filtration method to faecal coliform counts was tested on 88 water samples from the Port Harcourt area of the Rivers State of Nigeria. Seventy-eight streams/rivers and ten untreated distributive sources were analysed. High levels of suspended residues necessitated the filtration of 1-2 ml of the raw and 5-10 ml of diluted samples of the surface waters. Anaerobic incubation, overall, decreased the nonfaecal coliforms (NFCs) and faecal coliforms (FCs) by 69.32% and 48.86% respectively. Percent decrease of mean NFC and FC count in 21 streams sampled at random, was 8.85% and 20.88%, respectively. Results from a stream and a faecally polluted beach of a tidal river sampled periodically were variable. The NFCs and FCs decreases in the stream sample by 5.4% and 7.6% respectively were not significant. For the beach samples, the 9.8% reduction in NFCs was significant (p = 0.01), but the 4.19% increase in FCs was not. Seventy percent of the distributive samples gave zero FC counts. Anaerobic incubation reduced to zero FCs in 2 samples and NFCs in 5 samples. It is concluded that the advantages of anaerobic incubation are not commensurate with the increased expenditure which invariably is incurred in the routine analysis of these waters.

Bacteria, Anaerobic↗

PubMed2003

Human toxocariasis: a seroepidemiological survey in the municipality of Campinas (SP), Brazil.

The occurrence of human Toxocara infection was evaluated in three neighborhoods of the periphery of the Campinas municipality (Jardim Santa Mônica, Jardim São Marcos and Jardim Campineiro) in 1999. Forty residences and 138 residents were randomly selected by drawing lots and were submitted to a seroepidemiological survey, which included blood collection for the immunoenzymatic detection (ELISA) of anti-Toxocara antibodies and a blood count, and the application of a semi-structured questionnaire for the evaluation of epidemiological data. Significant levels of anti-Toxocara antibodies were detected in 23.9% of the 1999 samples. No significant difference in the frequency of infection according to age was observed. Environmental contamination with Toxocara eggs was observed in 12.3 and 14.0% of 57 soil samples collected in the same region in December 1998 and July 1999, respectively. Univariate analysis and multiple logistic regression of the data obtained from the questionnaires and of the results of the serological tests, suggest a significant influence of socioeconomic variables on the frequency of human infection with Toxocara under the conditions prevalent in the study area.

Adolescent↗

PubMed1983

Automated particle-counting immunoassay of C-reactive protein and its application to serum, cord serum, and cerebrospinal fluid samples.

The assay for C-reactive protein has been fully automated as a particle-counting immunoassay. For cerebrospinal fluid, cord serum, and adults' serum its range of sensitivity extends from 1 microgram/L to 300 mg/L, with a minimal sample dilution of twofold and a maximal dilution of 50-fold being required. This range is so broad because free antibodies are added to the reaction medium. However, we have used Fab fragments rather than whole antibody to avoid too steep a standard curve and a decrease of agglutination at high concentrations of antigen. For 99 consecutive cord sera examined, the concentrations of C-reactive protein ranged from 7 micrograms/L to 1.750 mg/L. The geometric mean was 50 micrograms/L and the upper normal limit (geometric mean +/- 2 SD of the log values) was established at 525 micrograms/L.

Adult↗

PubMed1983

[Application of an automatic blood cell counter, MICROX, to experimental animals. II. Leukocyte differential count in rabbits, rats, and mice].

Automatic blood cell analyzer "MICROX" was examined for possible applicability to the differential count of white blood cell in the rabbit, rat and mouse. Spun blood smear slides prepared from 37 rabbits were treated with Wright-Giemsa stain, and those from 49 rats and 46 mice were stained with Wright's dye. Totals of 4272 white blood cells of rabbit, 5484 cells of rat and 5329 cells of mouse were counted, respectively. The identification rate of MICROX was checked by the cell-by-cell counting method. Analysis of the data was made to examine correlation between the results with a MICROX and those of visual counting by optical microscopy on the same slides. 1) Corrected identification rates were 98.0%, 98.2% and 97.8% respectively in rabbits, rats and mice. 2) A high degree of correlation was observed between the results by MICROX and by optical microscopy in respect of segmented neutrophils and lymphocytes. 3) There were an average of 13.2 cells counted as unknown cells in rabbit blood, 10.5 in rat blood and 13.6 in mouse blood per 100 cells, respectively. Of the unknown cells about 30 to 40% were closely disposed leukocytes within the same counting squares. Almost all of atypical lymphocytes and erythroblasts were also counted as unknown cells on the analyzer. The data support the feasibility of the use of MICROX for the differential count of white blood cells of normal rabbits, rats and mice as well as monkeys and dogs.

Animals↗

PubMed2000

Measures of treatment effectiveness on tumour response and survival: a multi-state model approach.

This paper proposes the application of a counting process approach in the analysis of treatment effect on tumour response and survival. It relies on the definition of two transient states between which individuals may move over time, that is, response and non-response, and of one absorbing state, death. Three models are discussed according to the underlying Markov and duration dependence assumptions, as well as a marginal modelling of repeated transitions between the two transient states. We illustrate the proposed methods with data from a randomized clinical trial assessing the effect of fludarabine in patients with advanced chronic lymphocytic leukaemia.

Antineoplastic Agents↗

PubMed1983

A video-computer for the chronotropic and inotropic measurements of the beating of cultured heart cells.

A video-computer system has been developed to measure the chronotropy and inotropy of cultured heart cells. The motion of the cell is followed by recording of the changes of light intensity from the edge of a beating cell. With this system the velocity of contraction and relaxation, the time to peak contraction, relaxation time and the displacement of the cell in culture can be measured for the first time. Also, the rate of beating can be measured beyond the limits set by visual counting. With the application of this technique to the cultured heart cell system we have found that norepinephrine increases the velocity of relaxation thus reducing the ratio of contraction velocity to relaxation velocity, and reduces the twitch time. Increased external calcium, on the other hand, has little effect on either the velocity of contraction or relaxation but, like norepinephrine, decreases the twitch time.

Animals↗

PubMed2000

Biomass quantification by image analysis.

Microbiologists have always rely on microscopy to examine microorganisms. When microscopy, either optical or electron-based, is coupled to quantitative image analysis, the spectrum of potential applications is widened: counting, sizing, shape characterization, physiology assessment, analysis of visual texture, motility studies are now easily available for obtaining information on biomass. In this chapter the main tools used for cell visualization as well as the basic steps of image treatment are presented. General shape descriptors can be used to characterize the cell morphology, but special descriptors have been defined for filamentous microorganisms. Physiology assessment is often based on the use of fluorescent dyes. The quantitative analysis of visual texture is still limited in bioengineering but the characterization of the surface of microbial colonies may open new prospects, especially for cultures on solid substrates. In many occasions, the number of parameters extracted from images is so large that data-mining tools, such as Principal Components Analysis, are useful for summarizing the key pieces of information.

Algorithms↗

PubMed1995

Activation markers and cell proliferation as indicators of toxicity: a flow cytometric approach.

Cell proliferation is an attractive endpoint in in vitro toxicity assays, since nearly any kind of damage in a cell may result in altered cell proliferation. In toxicological applications, liquid scintillation counting, measuring radioactivity from tritiated thymidine, has been the traditional way to estimate cell proliferation. An alternative approach is the measurement of BrdU incorporation by flow cytometry. Before the actual DNA synthesis starts, several proteins are expressed on the cell surface, as well as intracellularly. Among the markers on the cell surface CD69, CD25, and CD71 are sequentially expressed on human lymphocytes after a mitogenic stimulation. The aim of this study was to evaluate information obtained by analysis of expression of activation markers on cell surfaces in lymphocyte subsets and to compare it with data from cell proliferation studies performed by liquid scintillation counting and BrdU flow cytometry. The experiments were performed with phytohemagglutinin-stimulated human lymphocytes exposed to ochratoxin A and cyclosporin A. While ochratoxin A-treated cultures showed a steep inhibition with increasing concentration, the cyclosporin A treatment gave an inhibition curve with a less steep slope. Activation marker studies showed that the effect of treatment with both of the toxins was more pronounced on the late markers CD25 and CD71, while CD69 had the advantage that significant effects could be detected as early as 6 h after ochratoxin A treatment. Cyclosporin A treatment induced only minor alterations in CD69 expression. Certain differences in expression of activation markers between CD4+ and CD8+ subsets were found both in ochratoxin A- and cyclosporin A-treated cultures. A stimulating effect was found in cell cultures exposed to the lowest concentration of ochratoxin A on CD69 and CD25 expression. Signs of an increase in frequencies of proliferating cells measured with the BrdU flow cytometry method were also seen. This increase could not be detected with liquid scintillation counting. No other differences between the liquid scintillation counting and BrdU flow cytometry measurements of proliferation were obtained. We conclude that studies of activation marker expression by the flow cytometric approach used in this report are useful complements to traditional measurements of cell proliferation as they yield subset-specific information about cellular processes which precede proliferation of lymphocytes.

Adult↗

PubMed2003

Measurement and prediction of METs during household activities in 35- to 45-year-old females.

This study determined whether four self-paced household tasks, conducted in the subjects' homes and a standardised laboratory environment, were performed at a moderate intensity [3-6 metabolic equivalents (METs)] in a representative sample of thirty-six 35- to 45-year-old females. Energy expenditure was also predicted via indirect methods. Self-paced energy expenditure during sweeping, window cleaning, vacuuming and mowing was measured using the Douglas bag technique. Heart rate, respiratory frequency, Computer Science Applications (CSA) movement counts (hip and wrist), Borg rating of perceived exertion and Quetelet's index were also recorded as potential predictors of energy expenditure. While the four activities were performed at mean intensities >or=3.0 METs in both the home and laboratory, all comparisons between these two environments were statistically significant ( P<0.001). The 95% confidence intervals (CIs) for the home and laboratory prediction equations were +/-1.1 METs and +/-1.0 MET, respectively. These data suggest that the aforementioned household chores can contribute to the 30 min x day(-1) of moderate-intensity activity required to confer health benefits. However, the substantial between-subject variability in energy expenditure resulted in some persons performing these tasks at a light intensity (<3.0 METs). The significant MET differences between the home and laboratory emphasise the effects of 'environment and terrain' and the 'mental approach to a task' on self-paced energy expenditure. Considering the means for the five activities ranged from 3.1 METs to 6.0 METs, the 95% CIs for the regression equations lack predictive precision.

Activities of Daily Living↗

PubMed1990

Random sampling by glitter drop method.

In the course of a study on glioblastoma tissue and cultured cell lines, a glitter drop technique for random sampling was introduced (1). Here, we report the initial description of the details of this procedure and its application to cell counts from explant slides and to determining bromodeoxyuridine (BUdR) labeling indices (LI) in tissues. The method is without bias, easy to perform and takes only a short time. It compares well with three other generally used random sampling procedures.

Bromodeoxyuridine↗

PubMed2005

Prostaglandin D2 and prostaglandin E2 accelerate the recovery of cutaneous barrier disruption induced by mechanical scratching in mice.

The role of prostaglandins in mechanical scratching-induced cutaneous barrier disruption in mice was investigated. Skin prostaglandins contents were measured after cutaneous barrier function was disrupted by scratching using a stainless-steal wire brush (mechanical scratching), then effects of prostanoids on recovery of cutaneous barrier functions were examined. This mechanical scratching increased transepidermal water loss and skin prostaglandins (prostaglandin D2, prostaglandin E2, 6-keto-prostaglandin F1alpha and prostaglandin F2alpha) contents, count-dependently. Topical application of indomethacin immediately after cutaneous barrier disruption delayed the recovery period of cutaneous barrier disruption. We examined effects of several prostanoids (prostaglandin D2, prostaglandin E2, prostaglandin F2alpha, prostaglandin I2 and U46619) on delay of the recovery process of mechanical scratching-induced cutaneous barrier disruption with treatment of indomethacin. Topically applied prostaglandin D2 and prostaglandin E2 accelerated the recovery of cutaneous barrier disruption and topical application of prostaglandin J2, limaprost, sulprostone and ONO-4819, but not 13,14-dihydro-15-keto-prostaglandin D2, 15-deoxy-Delta(12,14)-prostaglandin J2, 17-phenyl-trinor-prostaglandin E2 or butaprost had effects on recovery of the cutaneous barrier. These results suggest that prostaglandin D2 and prostaglandin E2 accelerate the recovery process of cutaneous barrier disruption caused by mechanical scratching, via specific prostanoid DP1, EP3 and EP4 receptors.

6-Ketoprostaglandin F1 alpha↗