Optimization and application of MTT assay in determining density of suspension cells.

MTT assay has been applied widely in stimulation and inhibition tests for adherent cells. However, reading step in regular MTT assays requires medium removal to dissolve formazan by absolute solvents results are usually affected by incomplete formazan dissolution and protein precipitation in diluted solvents. Therefore, most of current MTT assay procedures have difficulties in application on suspension cell lines. In this study, we demonstrated a modified MTT assay in which formazan dissolution method was improved by using the combination of DMSO and SDS-lysis solution. Importantly, the modified MTT assay did not require medium removal, thus it can be applicable for both suspension and adherent cell lines. We also verified that the modified MTT procedure could be effectively applied in bioactivity assays such as cancer cell inhibition and fibroblast stimulation assays. Besides the ease of use, our data regarding nonlinear regression model fitting, data variation, and separation clearly demonstrated that the sensitivity, stability and precision of modified assay were higher than those of common MTT procedures using isopropanol or DMSO as solvents. This study indicated that the modification described here can broaden the MTT assay application on suspension cell lines and also simplify the MTT protocol on adherent cell lines.

Impact of fibre intake and fibre source on digestibility, gut development, retention time and growth performance of indigenous and exotic pigs.

The impact of fibre level and fibre source on digestibility, gastrointestinal tract (GIT) development, total tract mean retention time (MRT) and growth performance was studied in indigenous Mong Cai (MC) and exotic Landrace × Yorkshire (LY) pigs. The diets were based on maize, rice bran, soyabean meal, fish meal and soyabean oil, and cassava residue (CR) or brewer's grain (BG) as fibrous ingredient sources in the high-fibre diets (HF) and were fed ad libitum. A low-fibre diet (LF), containing around 200 g NDF/kg dry matter (DM), was formulated without CR and BG as feed ingredients. The HF diets (HF-CR and HF-BG) were formulated to contain around 270 g NDF/kg DM. The experiment was arranged as a 2 × 3 factorial completely randomized design with six replications, and lasted 27 days. Increased dietary fibre level resulted in a reduction (P < 0.05) in average daily gain, digestibility of organic matter (OM), CP and gross energy (GE) at the ileum and in the total tract, and in MRT, and an increase (P < 0.05) in the feed conversion ratio and in the weight of the GIT (except for small intestine and caecum). The coefficients of total tract digestibility of fibre fractions were higher in HF diets than in the LF diet, with highest values for diet HF-CR, which had a high proportion of soluble non-starch polysaccharides. MC pigs had longer MRT of digesta than LY pigs (P < 0.05), resulting in higher digestibility at the ileum and in the total tract. Across diets and breeds, the total tract apparent digestibility of OM, CP and GE was positively related (R 2 = 0.80 to 0.84) to the MRT of solids, whereas the MRT was negatively related to the DM intake (R 2 = 0.60).

Effect of Fibre Level and Fibre Source on Gut Morphology and Micro-environment in Local (Mong Cai) and Exotic (Landrace×Yorkshire) Pigs.

The effect of genotype, fibre level and fibre source on gut morphology, environment and microflora was studied using 18 Mong Cai (MC) and 18 Landrace×Yorkshire (LY) pigs, aged around 60 d. The diets were based on maize, rice bran, soybean meal, fish meal and soybean oil, and cassava residue (CR) or brewer's grain (BG) as fibrous ingredient sources in the high-fibre diets (HF). A low-fibre diet (LF), containing around 200 g NDF/kg dry matter (DM), was formulated without CR and BG as feed ingredients. The HF diets (HF-CR and HF-BG) were formulated to contain around 270 g NDF/kg DM. The experiment was arranged according to a 2×3 factorial completely randomized design with six replications, and lasted 30 d. Crypt density in ileum was lowest (p<0.05) and villus height in jejunum and ileum were the greatest (p<0.05) in pigs fed diet HF-BG. Villus width in ileum was greatest in pigs fed diets HF-CR and HF-BG (p<0.05). Lactic acid bacteria (LAB) counts in stomach were greatest (p<0.05) and E. coli counts in ileum and colon were lowest (p<0.05) in pigs fed diet HF-CR. The concentration of total organic acids in ileum, caecum and colon were greatest (p<0.05), and pH in ileum and colon were lowest (p<0.05) in pigs fed diet HF-CR. Crypt density in ileum was lowest, and villus height in ileum and villus width in jejunum and ileum was greatest in LY pigs (p<0.05). LAB counts in stomach and ileum were greatest, and E. coli counts in ileum were lowest in MC pigs (p<0.05). The concentration of total organic acids in ileum, caecum and colon were greatest (p<0.05) and pH lowest (p<0.05) in MC pigs.

Chemical Characterization and Water Holding Capacity of Fibre-rich Feedstuffs Used for Pigs in Vietnam.

During two years, four samples per year were collected in Vietnam from rice bran, cassava residue, brewer's grain, tofu residue, soybean meal, coconut cake, sweet potato vines and water spinach for chemical analysis and assessment of water holding capacity (WHC). The selected feedstuffs represent fibre-rich plant sources and agro-industry co-products commonly used in pig feeding in Vietnam. The content (g/kg DM) of crude protein (CP), ether extract (EE) and non-starch polysaccharides (NSP) varied between feedstuffs and ranged from 21 to 506 for CP, from 14 to 118 for EE and from 197 to 572 for NSP. Cassava residue had a high starch content of 563 g/kg DM, while sweet potato vines, water spinach, coconut cake and soybean meal had a high content of sugars (63-71 g/kg DM). The content of individual neutral sugars varied between feed ingredients, with the highest content of arabinose, galactose and glucose in tofu residue, the highest content of xylose in brewer's grain and the highest content of mannose in coconut cake. The content of uronic acid was high for cassava residue, tofu residue, sweet potato vines and water spinach (57-88 g/kg DM). The content of soluble non-cellulosic polysaccharides (S-NCP) was positively correlated (r(2) = 0.82) to the WHC. The content (g/kg DM) of CP, NDF, neutral sugars, total NSP, total NCP, S-NCP and total dietary fibre in tofu residue, water spinach and coconut cake varied (p<0.05) between years. In conclusion, diet formulation to pigs can be improved if the variation in chemical composition of the fibre fraction and in WHC between potential feed ingredients is taken into account.