In this paper we report a quantitative laser Biospeckle method using VDRL plates to monitor the activity of Trypanosoma cruzi and the calibration conditions including three image processing algorithms and three programs (ImageJ and two programs designed in this work). Benznidazole was used as a test drug. Variable volume (constant density) and variable density (constant volume) were used for the quantitative evaluation of parasite activity in calibrated wells of the VDRL plate. The desiccation process within the well was monitored as a function of volume and of the activity of the Biospeckle pattern of the parasites as well as the quantitative effect of the surface parasite quantity (proportion of the object’s plane). A statistical analysis was performed with ANOVA, Tukey post hoc and Descriptive Statistics using R and R Commander. Conditions of volume (100μl) and parasite density (2-4x104 parasites/well, in exponential growth phase), assay time (up to 204min), frame number (11 frames), algorithm and program (RCommander/SAGA) for image processing were selected to test the effect of variable concentrations of benznidazole (0.0195 to 20μg/mL / 0.075 to 76.8μM) at various times (1, 61, 128 and 204min) on the activity of the Biospeckle pattern. The flat wells of the VDRL plate were found to be suitable for the quantitative calibration of the activity of Trypanosoma cruzi using the appropriate algorithm and program. Under these conditions, benznidazole produces at 1min an instantaneous effect on the activity of the Biospeckle pattern of T. cruzi, which remains with a similar profile up to 1 hour. A second effect which is dependent on concentrations above 1.25μg/mL and is statistically different from the effect at lower concentrations causes a decrease in the activity of the Biospeckle pattern. This effect is better detected after 1 hour of drug action. This behavior may be explained by an instantaneous effect on a membrane protein of Trypanosoma cruzi that could mediate the translocation of benznidazole. At longer times the effect may possibly be explained by the required transformation of the pro-drug into the active drug.

This paper reports on the application of the motion history image (MHI) method on dynamic laser speckle processing as a result of a specific drug action on Trypanosoma cruzi parasites. The MHI procedure is based on human action recognition, and unlike other methods which use a sequence consisting of several frames for recognition, this method uses only an MHI per action sequence for recognition. MHI method avoids the complexity as well as the large computation in sequence matching-based methods and detects a change in the speckle pattern. Experimental results of MHI on real-time monitoring of activity (motility) under the influence of the drug demonstrate the effectiveness of the proposed method. The MHI showed an online result without loss of resolution and definition if we compare with routine LASCA method. The obtained results highlight the advantage of the MHI analysis over traditional qualitative image intensity-based methods and demonstrate the potential of measuring the activity of parasites via dynamic laser speckle analysis. The data was further numerically analyzed in the time domain, and the results presented the ability of the technique to monitor the action of the drug, particularly Epirubicin (100 μg/ml).

This paper reports on the biospeckle processing of biological activity using a visualization scheme based upon the digital imaging information technology. Activity relative to bacterial growth in agar plates and to parasites affected by a drug is monitored via the speckle patterns generated by a coherent source incident on the microorganisms. We present experimental results to demonstrate the potential application of this methodology for following the activity in time. The digital imaging information technology is an alternative visualization enabling the study of speckle dynamics, which is correlated to the activity of bacteria and parasites. In this method, the changes in Red-Green-Blue (RGB) color component density are considered as markers of the growth of bacteria and parasites motility in presence of a drug. The RGB data was used to generate a two-dimensional surface plot allowing an analysis of color distribution on the speckle images. The proposed visualization is compared to the outcomes of the generalized differences and the temporal difference. A quantification of the activity is performed using a parameterization of the temporal difference method. The adopted digital image processing technique has been found suitable to monitor motility and morphological changes in the bacterial population over time and to detect and distinguish a short term drug action on parasites.

Disease mapping seeks to represent the risk of a disease. This paper focuses on the spatial analysis of risk for pandemic COVID-19 in Europe and the Mediterranean. Morbidity and mortality data for 54 countries in ratio format were used. Two hypotheses were considered, the first one is that the data are homogeneous and the second one is that the ratios are defined in a heterogeneous manner requiring the stratification on the basis of covariables and the methodology of Jenks’ intervals. Spatial risk models were applied as well as methods for the representation of clusters. The results show that the best representation is obtained with the Poisson-Gamma Model under stratification. The variations in the ratios are due to the individual policies of each country for the management of the pandemic. The cluster analysis shows that there is a high mortality process in Eastern Europe. The behavior of the pandemic should be evaluated in the space-time process as well as in other heterogeneous and highly unequal regions.

In this paper, we report on a biospeckle laser method used to monitor a specific drug action on T. cruzi parasites. Experimental results from fast biospeckle monitoring of the parasites' activity under the influence of the drug demonstrate the effectiveness of the proposed method. We measure the speckle parameters such as spatiotemporal correlation and speckle grain size to assess the immediate action of the drug on the parasites during a very short incubation period. From a practical point of view, this aproach allows us to validate biospeckle as a fast, non invasive and alternative method to test candidate drugs on T. cruzi parasites.