Reference 1D and 2D electrophoresis maps for potential disease related proteins in milk whey from lactating buffaloes and blood serum from buffalo calves (Water buffalo, Bubalus bubalis).

The aim of this study was to identify potential disease related proteins in milk whey of lactating buffaloes and blood serum of buffalo calves, in order to define a reference electrophoresis map for 1-DE and 2-DE. Additionally, changes in some protein patterns from buffalo calves during salmonellosis and lactating buffaloes during mastitis are presented. Milk samples were collected and distributed into groups: Milk samples from healthy buffaloes (SCC < 100.000 cells/ml, negative microbiology and CMT) (G1, n = 5) and buffaloes with subclinical mastitis (SCC > 500.000 cells/ml, positive microbiology and CMT) (G2, n = 5). Blood samples from buffalo calves (n = 6) were collected, and three calves were experimentally infected with Salmonella Dublin and samples analyzed before (M0) and 72 h after inoculation (M1). 1-DE was accomplished by loading 10 µg of TP into SDS-PAGE, stained with Coomassie blue. 2-DE was accomplished by loading 200 µg of TP into 11 cm, pH 3-10 non-linear IPG strips, followed by SDS-PAGE, stained with Coomassie blue. Protein bands/spots were excised, subjected to tryptic in-gel digestion and analyzed by LC/ESI-MS/MS. Protein identity was assigned using NCBI databases. After bands/spots from 1-DE and 2-DE were analyzed, a protein map with 35 and 40 different identified proteins in blood serum and milk whey, respectively, was generated. Significant changes in patterns of haptoglobin were observed in buffalo calves with salmonellosis and in patterns of IgLC, ß-lactoglobulin and α-lactalbumin of lactating buffaloes during mastitis. The establishment of a protein map for 1-DE and 2-DE, identifying potential disease related proteins, can help to address alterations during diseases in buffaloes.

Pseudomonas spp: contamination sources in bulk tanks of dairy farms / Pseudomonas spp: fontes de contaminação em tanques de expansão em fazendas leiteiras

This study focused on isolating Pseudomonas spp. during milking process in ten dairy farms with manual and mechanical milking systems during dry and rainy seasons, and evaluating DNA homology and patterns of distribution between isolates, in order to identify main sources of milk contamination by Pseudomonas spp. A total of 167 isolates of Pseudomonas spp. were obtained from water, milkers' hands, cows' teats, teat cups, cooling tanks and raw milk. Bacteria of Pseudomonas spp. genus were isolated from 85 and 82 sampling points in dairy farms with manual and mechanical milking system, respectively. A significant difference (p=0.02) on Pseudomonas spp. isolation was observed among samples of surface of cows' teats before and after pre-dipping, but no significant difference (p>0.05) was observed among milking systems or seasons. The possibility of the same Pseudomonas spp. patterns are distributed in different farms and seasons using Amplified Fragment Length Polymorphism (AFLP) technique was demonstrated. Milkers' hands, surface of cows' teats, teat cups and cooling tanks were associated with raw milk contamination with Pseudomonas spp. on farms with manual and mechanical milking system, showing that regardless of the type of milking system and season, proper hygiene procedures of equipment, utensils and workers' hands are essential to avoid contamination of the milk and, therefore, improve milk quality.(AU)

Este estudo se propôs a isolar Pseudomonas spp. durante o processo de ordenha em dez fazendas com sistemas manuais e mecanizados, durante as estações seca e chuvosa, além de avaliar a homologia do DNA e seus padrões de distribuição entre os isolados, a fim de se determinar as principais fontes de contaminação do leite. Cento e sessenta e sete isolados de Pseudomonas spp. foram obtidos a partir de amostras de água, mãos de ordenhadores, tetos, teteiras, tanques de resfriamento e leite cru armazenado, sendo 85 e 82 pontos de amostragem em fazendas com sistemas de ordenha manual e mecânico, respectivamente. Diferença estatisticamente significativa foi encontrada entre os isolados observados entre a superfície dos tetos antes e após o pré-dipping (p=0,02), mas nenhuma diferença foi encontrada entre sistemas de ordenha ou estações (p>0,05). A possibilidade do mesmo padrão de Pseudomonas spp. estar distribuído em diferentes fazendas ou estações foi avaliada pela técnica de Polimorfismo do Tamanho de Fragmento Amplificado (AFLP). As mãos de ordenhadores, superfície dos tetos das vacas, teteiras e tanques de resfriamento foram associados com a contaminação do leite cru, demonstrando que independente do tipo de ordenha e estação, a higiene adequada de equipamentos, utensílios e mãos dos ordenhadores é essencial para evitar contaminação do leite, e consequentemente aumentar sua qualidade.(AU)