Biological activity using dynamic speckle in blood serum of mares in different reproductive stages / Atividade biológica por speckle dinâmico em soro sanguíneo de éguas em diferentes estágios reprodutivos

This study aimed to evaluate the biological activity in blood serum plasma samples from mares at different reproductive stages using dynamic speckle analysis. For this purpose, samples were collected from 40 Mangalarga Machador mares from the Formoso 2S horse farm in Cajueiro, AL. The mares were classified into four groups of ten animals each according to the pregnancy stage: ten empty mares (not pregnant), ten pregnant mares in the early third of pregnancy between one month and four months, ten pregnant mares in the final third of the pregnancy between seven and eleven months, and ten lactating mares (within two months post-partum). The biological activity response using a dynamic speckle (biospeckle) was obtained by capturing images reflected by a coherent light (diode laser with a wavelength of 532 nm and intensity of 3 mW) on a sample using a CCD (charge coupled device) camera. This data was analyzed using image processing techniques through the computational application SpeckleTHSP-MCO-Descritores (STMD), applying THSP (time history speckle pattern) methodology, which evaluates the temporal evolution of the interference image from spreading over the sample surface over time. A coherence matrix generated from the THSP was used to present the intensity module dispersion, which provided the bioactivity data. These data were then processed using the program OriginPro 8. Ink, generate graphs and compare the results from the different groups under study. A general biospeckle signature was observed regardless of the mares reproductive stage considered in this study. A short phase of movement of the samples associated with an accommodation of the drop on the slide was observed. It was followed by an ascending curve starting between 5 and 10 min of observation, reaching a peak within 15 and 25 min, and finally decayed uniformly until almost zero after 45 min. The group of pregnant mares in the final third of pregnancy presented superior bioactivity compared to pregnant mares in the early third of pregnancy. The curve observed for the group of lactating mares is similar to the curve obtained for the group of pregnant mares in the early third of pregnancy. Bioactive molecules act as dispersion elements of coherent light incident on a sample. The variation inherent to the presence of bioactive molecules in the different stages evaluated influenced the biospeckle pattern detected in each sample. It was concluded that the biological activity peaks of the blood plasma samples from the mares evaluated in this study using the dynamic speckle analysis technique were different for both amplitude and time of occurrence, according to the different reproductive stages.(AU)

Objetivou-se com esse estudo, avaliar os picos de atividade biológica de amostras de plasma sanguíneo de éguas em diferentes estágios reprodutivos, por análise de speckle dinâmico. Foram utilizadas amostras de 40 éguas da raça Mangalarga Marchador pertencentes ao Haras Formoso 2S localizado na cidade de Cajueiro AL. As éguas foram divididas em 4 grupos de 10 animais cada, classificados de acordo com a fase de gestação, sendo 10 éguas vazias (não prenhes), 10 éguas prenhas no terço inicial de gestação entre 1 e 4 meses, 10 éguas em terço final de gestação entre 7 e 11 meses e 10 éguas em fase de lactação (até o segundo mês pós-parto). A obtenção da leitura de atividade biológica por speckle dinâmico (biospeckle) foi feita através da captura de imagens refletidas por uma luz coerente (laser de diodo com comprimento de onda de 532 nm e intensidade de 3mW) por meio de uma câmera CCD (charge coupled device) nas amostras de soro sanguíneo. Esses dados foram analisados através de técnicas de processamento de imagens pelo aplicativo computacional Speckle-THSP-MCO-Descritores (STMD), empregando-se o método de THSP (time history speckle pattern), que avalia a evolução temporal da figura de interferência a partir do espalhamento pela superfície da amostra ao longo do tempo. A partir do THSP foi gerada uma matriz de co-ocorrência, usada para apresentar o MDI (módulo de dispersão de intensidades), que forneceu os dados da bioatividade das amostras. Os dados obtidos foram então tratados pelo programa OriginPro 8.Ink, para geração de gráficos e comparação dos mesmos entre os grupos estudados. Observou-se que, independente do estágio reprodutivo das éguas considerado nesse estudo, existe uma assinatura geral de atividade captada pelo biospeckle. Ocorreu uma curta fase de movimentação das amostras, provavelmente associada à acomodação da gota na lâmina, seguida de uma curva ascendente iniciada entre o minuto 5 e 10 de avaliação, que alcança seu pico em uma faixa de tempo entre o minuto 15 e 25, decaindo de maneira uniforme até praticamente se anular quando passados 45 minutos de análise. O grupo de éguas em terço final de gestação apresentou bioatividade das amostras superior, quando comparadas às éguas que estão em terço inicial. A curva observada no grupo de éguas em lactação se assemelha um pouco à curva do grupo de terço inicial. A variação inerente à presença de moléculas bioativas nas diferentes fases avaliadas, que atuam como elementos dispersores de uma luz coerente incidida nas amostras, aparentemente influenciam no padrão de biospeckle detectado em cada uma delas. Conclui-se que os picos de atividade biológica de amostras de plasma sanguíneo de éguas avaliadas pela técnica de análise de speckle dinâmico, mostram-se distintos tanto em amplitude quanto em tempo de ocorrência, de acordo com os estágios reprodutivos das fêmeas.(AU)

Biological activity using dynamic speckle in blood serum of mares in different reproductive stages / Atividade biológica por speckle dinâmico em soro sanguíneo de éguas em diferentes estágios reprodutivos

This study aimed to evaluate the biological activity in blood serum plasma samples from mares at different reproductive stages using dynamic speckle analysis. For this purpose, samples were collected from 40 Mangalarga Machador mares from the Formoso 2S horse farm in Cajueiro, AL. The mares were classified into four groups of ten animals each according to the pregnancy stage: ten empty mares (not pregnant), ten pregnant mares in the early third of pregnancy between one month and four months, ten pregnant mares in the final third of the pregnancy between seven and eleven months, and ten lactating mares (within two months post-partum). The biological activity response using a dynamic speckle (biospeckle) was obtained by capturing images reflected by a coherent light (diode laser with a wavelength of 532 nm and intensity of 3 mW) on a sample using a CCD (charge coupled device) camera. This data was analyzed using image processing techniques through the computational application SpeckleTHSP-MCO-Descritores (STMD), applying THSP (time history speckle pattern) methodology, which evaluates the temporal evolution of the interference image from spreading over the sample surface over time. A coherence matrix generated from the THSP was used to present the intensity module dispersion, which provided the bioactivity data. These data were then processed using the program OriginPro 8. Ink, generate graphs and compare the results from the different groups under study. A general biospeckle signature was observed regardless of the mares’ reproductive stage considered in this study. A short phase of movement of the samples associated with an accommodation of the drop on the slide was observed. It was followed by an ascending curve starting between 5 and 10 min of observation, reaching a peak within 15 and 25 min, and finally decayed uniformly until almost zero after 45 min. The group of pregnant mares in the final third of pregnancy presented superior bioactivity compared to pregnant mares in the early third of pregnancy. The curve observed for the group of lactating mares is similar to the curve obtained for the group of pregnant mares in the early third of pregnancy. Bioactive molecules act as dispersion elements of coherent light incident on a sample. The variation inherent to the presence of bioactive molecules in the different stages evaluated influenced the biospeckle pattern detected in each sample. It was concluded that the biological activity peaks of the blood plasma samples from the mares evaluated in this study using the dynamic speckle analysis technique were different for both amplitude and time of occurrence, according to the different reproductive stages.

Objetivou-se com esse estudo, avaliar os picos de atividade biológica de amostras de plasma sanguíneo de éguas em diferentes estágios reprodutivos, por análise de speckle dinâmico. Foram utilizadas amostras de 40 éguas da raça Mangalarga Marchador pertencentes ao Haras Formoso 2S localizado na cidade de Cajueiro – AL. As éguas foram divididas em 4 grupos de 10 animais cada, classificados de acordo com a fase de gestação, sendo 10 éguas vazias (não prenhes), 10 éguas prenhas no terço inicial de gestação entre 1 e 4 meses, 10 éguas em terço final de gestação entre 7 e 11 meses e 10 éguas em fase de lactação (até o segundo mês pós-parto). A obtenção da leitura de atividade biológica por speckle dinâmico (biospeckle) foi feita através da captura de imagens refletidas por uma luz coerente (laser de diodo com comprimento de onda de 532 nm e intensidade de 3mW) por meio de uma câmera CCD (charge coupled device) nas amostras de soro sanguíneo. Esses dados foram analisados através de técnicas de processamento de imagens pelo aplicativo computacional Speckle-THSP-MCO-Descritores (STMD), empregando-se o método de THSP (time history speckle pattern), que avalia a evolução temporal da figura de interferência a partir do espalhamento pela superfície da amostra ao longo do tempo. A partir do THSP foi gerada uma matriz de co-ocorrência, usada para apresentar o MDI (módulo de dispersão de intensidades), que forneceu os dados da bioatividade das amostras. Os dados obtidos foram então tratados pelo programa OriginPro 8.Ink, para geração de gráficos e comparação dos mesmos entre os grupos estudados. Observou-se que, independente do estágio reprodutivo das éguas considerado nesse estudo, existe uma assinatura geral de atividade captada pelo biospeckle. Ocorreu uma curta fase de movimentação das amostras, provavelmente associada à acomodação da gota na lâmina, seguida de uma curva ascendente iniciada entre o minuto 5 e 10 de avaliação, que alcança seu pico em uma faixa de tempo entre o minuto 15 e 25, decaindo de maneira uniforme até praticamente se anular quando passados 45 minutos de análise. O grupo de éguas em terço final de gestação apresentou bioatividade das amostras superior, quando comparadas às éguas que estão em terço inicial. A curva observada no grupo de éguas em lactação se assemelha um pouco à curva do grupo de terço inicial. A variação inerente à presença de moléculas bioativas nas diferentes fases avaliadas, que atuam como elementos dispersores de uma luz coerente incidida nas amostras, aparentemente influenciam no padrão de biospeckle detectado em cada uma delas. Conclui-se que os picos de atividade biológica de amostras de plasma sanguíneo de éguas avaliadas pela técnica de análise de speckle dinâmico, mostram-se distintos tanto em amplitude quanto em tempo de ocorrência, de acordo com os estágios reprodutivos das fêmeas.

Fermentation process for production of apple-based kefir vinegar: microbiological, chemical and sensory analysis

The aim of this study was to develop a kefir apple-based vinegar and evaluate this fermentation process using new methodology with Biospeckle Laser. Brazilian kefir grains were inoculated in apple must for vinegar production. In this study, the microbial community present in kefir, and correspondent vinegar, was investigated using Matrix Assisted Laser Desorption/Ionization - Time of Flight Mass Spectrometry (MALDI-TOF MS) technique. Saccharomyces cerevisiae, Lactobacillus paracasei, Lactobacillus plantarum, Acetobacter pasteurianus and Acetobacter syzygii were the microbial species identified. S. cerevisiae, L. plantarum, A. pasteurianus and A. syzygii were found in smaller quantities at the beginning of the alcoholic fermentation, but were found throughout the alcoholic and acetic fermentation. Kefir grains were able to utilize apple must as substrate to produce ethanol, and acetic acid. Acetate, volatile alcohols and aldehydes in the vinegar-based kefir were also produced. The yield of acetic acid in the kefir vinegars was ∼79%. The acetic acid concentration was ∼41 g L-1, reaching the required standard for the Brazilian legislation accepts it as vinegar (4.0% acetic acid). Kefir vinegar showed good acceptance in the sensory analysis. The technology proposed here is novel by the application of immobilized-cell biomass (kefir grains) providing a mixed inocula and eliminating the use of centrifuge at the end of the fermentative process. This step will save energy demand and investment. This is the first study to produce apple vinegar using kefir grains.(AU)

Fermentation process for production of apple-based kefir vinegar: microbiological, chemical and sensory analysis

Abstract The aim of this study was to develop a kefir apple-based vinegar and evaluate this fermentation process using new methodology with Biospeckle Laser. Brazilian kefir grains were inoculated in apple must for vinegar production. In this study, the microbial community present in kefir, and correspondent vinegar, was investigated using Matrix Assisted Laser Desorption/Ionization - Time of Flight Mass Spectrometry (MALDI-TOF MS) technique. Saccharomyces cerevisiae, Lactobacillus paracasei, Lactobacillus plantarum, Acetobacter pasteurianus and Acetobacter syzygii were the microbial species identified. S. cerevisiae, L. plantarum, A. pasteurianus and A. syzygii were found in smaller quantities at the beginning of the alcoholic fermentation, but were found throughout the alcoholic and acetic fermentation. Kefir grains were able to utilize apple must as substrate to produce ethanol, and acetic acid. Acetate, volatile alcohols and aldehydes in the vinegar-based kefir were also produced. The yield of acetic acid in the kefir vinegars was ∼79%. The acetic acid concentration was ∼41 g L-1, reaching the required standard for the Brazilian legislation accepts it as vinegar (4.0% acetic acid). Kefir vinegar showed good acceptance in the sensory analysis. The technology proposed here is novel by the application of immobilized-cell biomass (kefir grains) providing a mixed inocula and eliminating the use of centrifuge at the end of the fermentative process. This step will save energy demand and investment. This is the first study to produce apple vinegar using kefir grains.

Digital imaging information technology for biospeckle activity assessment relative to bacteria and parasites.

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.

Fermentation process for production of apple-based kefir vinegar: microbiological, chemical and sensory analysis.

The aim of this study was to develop a kefir apple-based vinegar and evaluate this fermentation process using new methodology with Biospeckle Laser. Brazilian kefir grains were inoculated in apple must for vinegar production. In this study, the microbial community present in kefir, and correspondent vinegar, was investigated using Matrix Assisted Laser Desorption/Ionization - Time of Flight Mass Spectrometry (MALDI-TOF MS) technique. Saccharomyces cerevisiae, Lactobacillus paracasei, Lactobacillus plantarum, Acetobacter pasteurianus and Acetobacter syzygii were the microbial species identified. S. cerevisiae, L. plantarum, A. pasteurianus and A. syzygii were found in smaller quantities at the beginning of the alcoholic fermentation, but were found throughout the alcoholic and acetic fermentation. Kefir grains were able to utilize apple must as substrate to produce ethanol, and acetic acid. Acetate, volatile alcohols and aldehydes in the vinegar-based kefir were also produced. The yield of acetic acid in the kefir vinegars was ∼79%. The acetic acid concentration was ∼41gL-1, reaching the required standard for the Brazilian legislation accepts it as vinegar (4.0% acetic acid). Kefir vinegar showed good acceptance in the sensory analysis. The technology proposed here is novel by the application of immobilized-cell biomass (kefir grains) providing a mixed inocula and eliminating the use of centrifuge at the end of the fermentative process. This step will save energy demand and investment. This is the first study to produce apple vinegar using kefir grains.

Continuous, high-resolution biospeckle imaging reveals a discrete zone of activity at the root apex that responds to contact with obstacles.

BACKGROUND AND AIMS: Shining a laser onto biological material produces light speckles termed biospeckles. Patterns of biospeckle activity reflect changes in cell biochemistry, developmental processes and responses to the environment. The aim of this work was to develop methods to investigate the biospeckle activity in roots and to characterize the distribution of its intensity and response to thigmostimuli. METHODS: Biospeckle activity in roots of Zea mays, and also Jatropha curcas and Citrus limonia, was imaged live and in situ using a portable laser and a digital microscope with a spatial resolution of 10 µm per pixel and the ability to capture images every 0.080 s. A procedure incorporating a Fujii algorithm, image restoration using median and Gaussian filters, image segmentation using maximum-entropy threshold methods and the extraction of features using a tracing algorithm followed by spline fitting were developed to obtain quantitative information from images of biospeckle activity. A wavelet transform algorithm was used for spectral decomposition of biospeckle activity and generalized additive models were used to attribute statistical significance to changes in patterns of biospeckle activity. KEY RESULTS: The intensity of biospeckle activity was greatest close to the root apex. Higher frequencies (3-6 Hz) contributed most to the total intensity of biospeckle activity. When a root encountered an obstacle, the intensity of biospeckle activity decreased abruptly throughout the root system. The response became attenuated with repeated thigmostimuli. CONCLUSIONS: The data suggest that at least one component of root biospeckle activity resulted from a biological process, which is located in the zone of cell division and responds to thigmostimuli. However, neither individual cell division events nor root elongation is likely to be responsible for the patterns of biospeckle activity.