Separation and quantification of milk proteins with the addition of cheese whey by lab-on-a-chip

O objetivo deste trabalho foi avaliar a eletroforese com dispositivo de microfluidos, conhecida como técnica lab-on-a-chip, para detecção de adulteração de leite com soro de queijo, em comparação ao SDS-PAGE. Amostras de leite cru, pasteurizado, processado em temperatura ultra-alta (UHT) e em pó receberam adição de soro de queijo em concentrações crescentes (0, 1, 2,5, 5, 10, 20, 30, 50 e 100% v/v) e foram submetidas a eletroforese lab-on-a-chip e SDS-PAGE para detectar suas misturas. A metodologia lab-on-a-chip foi capaz de separar e quantificar as proteínas do leite. Além disso, a técnica lab-on-a-chip é fácil, rápida, sensível e pode detectar adição de soro de queijo no leite do nível mais baixo testado (1%) para as proteínas do leite α-caseína e β-caseína.The objective of this work was to evaluate microfluidic chip electrophoresis, known as lab-on-a-chip technique, for the detection of milk adulteration using cheese whey in comparison with SDS-PAGE. Raw, pasteurized, processed at an ultra-high temperature (UHT), and powdered milk samples received increasing concentrations of cheese whey (0, 1, 2.5, 5, 10, 20, 30, 50, and 100% v/v), and were subjected to lab-on-a-chip electrophoresis and SDS-PAGE to detect their mixtures. The lab-on-a-chip methodology was able to separate and quantify milk proteins. In addition, the tested technique is easy, rapid, sensitive, and can detect the addition of cheese whey in milk from the lowest level tested (1%) for milk proteins α-casein and β-casein

Expression profile of genes associated with mastitis in dairy cattle

In order to characterize the expression of genes associated with immune response mechanisms to mastitis, we quantified the relative expression of the IL-2, IL-4, IL-6, IL-8, IL-10, IFN-γ and TNF- α genes in milk cells of healthy cows and cows with clinical mastitis. Total RNA was extracted from milk cells of six Black and White Holstein (BW) cows and six Gyr cows, including three animals with and three without mastitis per breed. Gene expression was analyzed by real-time PCR. IL-10 gene expression was higher in the group of BW and Gyr cows with mastitis compared to animals free of infection from both breeds (p < 0.05). It was also higher in BW Holstein animals with clinical mastitis (p < 0.001), but it was not significant when Gyr cows with and without mastitis were compared (0.05 < p < 0.10). Among healthy cows, BW Holstein animals tended to present a higher expression of all genes studied, with a significant difference for the IL-2 and IFN- γ genes (p < 0.001). For animals with mastitis no significant difference in gene expression was observed between the two breeds. These findings suggest that animals with mastitis develop a preferentially cell-mediated immune response. Further studies including larger samples are necessary to better characterize the gene expression profile in cows with mastitis

Candidate gene expression in bos indicus ovarian tissues: prepubertal and postpubertal heifers in diestrus

Growth factors such as bone morphogenetic proteins 6, 7, 15, and two isoforms of transforming growth factor-beta (BMP6, BMP7, BMP15, TGFB1, and TGFB2), and insulin-like growth factor system act as local regulators of ovarian follicular development. To elucidate if these factors as well as others candidate genes, such as estrogen recep- tor 1 (ESR1), growth differentiation factor 9 (GDF9), follicle-stimulating hormone receptor (FSHR), luteinizing hormone receptor (LHR), bone morphogenetic protein receptor, type 2 (BMPR2), type 1 insulin-like growth factor receptor (IGFR1), and key steroidogenic enzymes cytochrome P450 aromatase and 3-β-hydroxysteroid dehydrogenase (CYP19A1 and HSD3B1) could modulate or influence diestrus on the onset of puberty in Brahman heifers, their ovarian mRNA expression was measured before and after puberty (luteal phase). Six postpubertal (POST) heifers were euthanized on the luteal phase of their second cycle, confirmed by corpus luteum observation, and six prepubertal (PRE) heifers were euthanized in the same day. Quantitative real-time PCR analysis showed that the expression of FSHR, BMP7, CYP19A1, IGF1, and IGFR1 mRNA was greater in PRE heifers, when contrasted to POST heifers. The expression of LHR and HSD3B1 was lower in PRE heifers. Differential expression of ovarian genes could be associated with changes in follicular dynamics and different cell populations that have emerged as consequence of puberty and the luteal phase. The emerging hypothesis is that BMP7 and IGF1 are co-expressed and may modulate the expression of FSHR, LHR and IGFR1, and CYP19A1. BMP7 could influence the downregulation of LHR and upregulation of FSHR and CYP19A1, which mediates the follicular dynamics in heifer ovaries. Upregulation of IGF1 expression prepuberty, compared to postpuberty diestrus, correlates with increased levels FSHR and CYP19A1. Thus, BMP7 and IGF1 may play synergic roles and were predicted to interact, from the expression data (P = 0.07, r = 0.84). The role of these co-expressed genes in puberty and heifers luteal phase merits further research