342 research outputs found

    Influence of Listeria innocua on the growth of Listeria monocytogenes

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    The growth of Listeria monocytogenes and Listeria innocua strains was monitored during this study: (i) in TSB–YE media and (ii) in a food matrix (pasteurized milk) according to the ISO 11290-1 methodology. Different inocula concentrations and mixtures were tested. The response was shown to be strain dependent. In TSB–YE the inhibition of a L. monocytogenes strain was observed in just one of the three mixtures (L. monocytogenes_1340 with L. innocua_11288) showing a reduction of 1.37 log cfu/ml after 42.5 h and 1.85 log after 66.5 h of incubation. In pasteurized milk the inhibition of L. monocytogenes by L. innocua was always observed when L. innocua was present in higher concentrations than L. monocytogenes. The reverse was also observed but only in one mixture (cocktail of six L. monocytogenes with L. innocua_ 2030c) when the initial concentration of L. monocytogenes was 100 times higher than L. innocua suggesting the phenomenon of quorum sensing. Furthermore, inhibitory activity was not caused by bacteriocins, and no correlation between the growth rate and inhibition was demonstrated.info:eu-repo/semantics/acceptedVersio

    Intracellular poly-P assessment by DAPI staining and image analysis

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    In wastewater treatment, enhanced biological phosphorus removal (EBPR) is considered a well-established process to remove phosphate (P). EBPR is based on the activity of polyphosphate-accumulating organisms (PAOs) able to take up and store large amounts of P as intracellular (poly-P) granules. However, monitoring poly-P in mixed cultures is usually performed by a laborious and time consuming off-line chemical analysis. Thus, there is a clear need to develop new techniques to rapidly monitor these processes, such as image analysis coupled to sample staining and microscopy inspection. A lab-scale sequencing batch reactor (SBR) was fed with synthetic wastewater containing acetate and propionate as main carbon sources and an orthophosphate solution was added. A COD/P ratio of 10 mg COD mg P-PO4-1 was used to provide selective advantages to PAOs. The SBR was operated with a cycle time of 6 h: 120 min anaerobic including 5 min feed, 180 min aerobic and 60 min wasting/settling. Biomass samples were collected at the end of the aerobic stage. Bulk P concentration was determined by segmented flow analysis and total P concentration was similarly measured following acid digestion at 100oC. Intracellular poly-P concentration was determined by subtracting the bulk P from the total P. Intracellular poly-P granules were observed in epifluorescence microscopy using DAPI staining with a 25 ìg mL-1 DAPI solution. A long pass filter was used with an excitation bandpass of 365-370 nm and emission cut off at 421 nm. A specially developed program in Matlab was used for image analysis. A total of 41 samples were collected. Two thirds were fed as training data to the partial least squares (PLS) model and the remaining used for validation. Both absolute (in mg poly-P / L) and relative (in mg poly-P / g MLSS) intracellular poly-P concentrations were studied. This procedure was found to predict, at some extent, the relative intracellular poly-P concentration (real poly-P = 0.971 x predicted poly-P, R2 of 0.744). Regarding the absolute intracellular poly-P concentration, a total of 3 samples needed to be discarded in order to obtain a similar result (real poly-P = 1.005 x predicted poly-P, R2 of 0.731)

    Canine parvovirus 2c infection in central Portugal

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    Canine parvovirus (CPV) has been evolving, generating new genetic and antigenic variants throughout the world. This Study was conducted to determine the types of CPV circulating in dogs in Figueira da Foz, Portugal. Thirty fecal samples, collected between 2006 and 2007 from dogs with clinical signs of CPV infection, were tested for CPV by a rapid, in-clinic, enzyme-linked immunosorbent assay (ELISA)/immunomigration test, by conventional real-time polymerase chain reaction (PCR), and by minor-groove binding TaqMan PCR. Of the 29 PCR-positive samples, 15 were identified as CPV-2b and 14 as CPV-2c. No CPV-2a was detected. The sensitivity of the ELISA test was 82.76% compared with the PCR assays. No significant associations were found between CPV type, clinical outcome, breed, vaccination status, or age

    Novel method to quantify intracellular accumulation of polyphosphate in EBPR systems

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    A new method for intracellular storage polyphosphate (poly-P)identification and quantification in enhanced biological phosphorus removal (EBPR) systems is proposed based on image analysis. In EBPR systems, 4',6-diamidino-2-phenylindole (DAPI) is usually combined with fluororescent in situ hybridization (FISH) to evaluate the microbial community. The proposed technique is based on an image analysis procedure specifically developed for determining poly-P inclusions within biomass suspension using solely DAPI by epifluorescence microscopy. Due to contradictory literature DAPI concentrations used for poly-P detection, the present work assessed the optimal DAPI concentration for samples acquired at the end of the EBPR aerobic stage when the accumulation is performed. Digital images were then acquired and processed by means of image processing and analysis. Regarding image analysis results and considering the current operational conditions, a promising correlation could be found between average poly-P intensity values and the analytical determination, although presenting a correlation coefficient somewhat far from the ideal. The proposed methodology can be seen as a promising alternative procedure to quantify intracellular poly-P accumulation in a faster and less labor intensive way

    Biopolymer monitoring using quantitative image analysis techniques

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    Polyhydroxyalkanoates (PHAs) are intracellular biopolymers with many applications, particularly as substitutes of polypropylene and polyethylene, due to their thermoplastic properties and biocompatible nature. Furthermore, glycogen is a polysaccharide of glucose with high importance in the metabolism of microbial communities and polyphosphate is a microbial storage compound that should be recovered in order to offset the worldwide depletion of phosphorus sources. The determination of these biopolymers by chemical analysis is a laborious task, often involving digestion processes prior to gas and high-performance liquid chromatography, which are time consuming and difficult to apply in industry. Currently, it is important to develop new, rapid and simple techniques to monitor these polymers. Image analysis is a non-invasive and rapid technique that has the potential to be used to quantify these intracellular polymers quickly, in real-time. Mesquita et al. (2013) showed that it is possible to predict the concentration of glycogen and PHAs by quantitative image analysis, using aniline blue and nile blue staining, respectively. Polyphosphate can also be predicted by this technique through DAPI staining, which is currently under development. These biopolymers are produced by several different microorganisms, and combining their quantification with fluorescence in situ hybridization (FISH) techniques for microbial identification can enable the determination of organisms that store high quantities of each biopolymer. In this work, an advanced quantitative technique is developed to perform real time monitoring of these three biopolymers in a bioreactor performing biological phosphorus removal. Image analysis of the biopolymers was combined with FISH to determine the storage level of each compound within the different microbial populations. This technique will further enable the assessment of biopolymer levels within microbial communities, which can be applied in the biopolymer production industry

    An Eye for Possibilities in the Development of Children with Cerebral Palsy: Neurobiology and Neuropsychology in a Cultural-Historical Dynamic Understanding

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    Taking children with Cerebral Palsy (CP) as an example, the article seeks an understanding ofchildren with disabilities that connects neuropsychological theories of neural development withthe situated cognition perspective and the child as an active participant in its social practices. Theearly brain lesion of CP is reconceptualised as a neurobiological constraint that exists in therelations between the neural, cognitive and social levels. Through a multi-method study of twochildren with CP, it is analysed how neurobiological constraints arise, evolve and sometimes areresolved through local matches between the child and its social practices. The result is discussedas support of a developmental science approach that includes processes at the social practice levelalong with knowledge of biological processes

    Genotype imputation strategies for Portuguese Holstein cattle using different SNP panels.

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    Abstract: Although several studies have investigated the factors affecting imputation accuracy, most of these studies involved a large number of genotyped animals. Thus, results from these studies cannot be directly applied to small populations, since the population structure affects imputation accuracy. In addition, factors affecting imputation accuracy may also be intensified in small populations. Therefore, we aimed to compare different imputation strate-gies for the Portuguese Holstein cattle population considering several commercially available single nucleotide poly-morphism (SNP) panels in a relatively small number of genotyped animals. Data from 1359 genotyped animals were used to evaluate imputation in 7 different scenarios. In the S1 to S6 scenarios, imputations were performed from LDv1, 50Kv1, 57K, 77K, HDv3 and Ax58K panels to 50Kv2 panel. In these scenarios, the bulls in 50Kv2 were divided into reference (352) and validation (101) populations based on the year of birth. In the S7 scenario, the validation population consisted of 566 cows genotyped with the Ax58K panel with theirgenotypes masked to LDv1. In general, all sample imputation accuracies were high with correlations ranging from 0.94 to 0.99 and concordance rate rang-ing from 92.59 to 98.18%. SNP-specific accuracy was consistent with that of sample imputation. S4 (40.32% of SNPs imputed) had higher accuracy than S2 and S3, both with less than 7.59% of SNPs imputed. Most probably, this was due to the high number of imputed SNPs with minor allele frequency (MAF) < 0.05 in S2 and S3 (by 18.43% and 16.06% higher than in S4, respectively). Therefore, for these two scenarios, MAF was more relevant than the panel density. These results suggest that genotype imputation using several commercially available SNP panels is feasible for the Portuguese national genomic evaluation
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