Comparison of methods for analysis of proteolysis by plasmin in milk

Sensitive methods that are currently used to monitor proteolysis by plasmin in milk are limited due to 7 their high cost and lack of standardisation for quality assurance in the various dairy laboratories. In 8 this study, four methods, trinitrobenzene sulphonic acid (TNBS), reverse phase high pressure liquid 9 chromatography (RP-HPLC), gel electrophoresis and fluorescamine, were selected to assess their 10 suitability for the detection of proteolysis in milk by plasmin. Commercial UHT milk was incubated 11 with plasmin at 37 °C for one week. Clarification was achieved by isoelectric precipitation (pH 4·6 12 soluble extracts)or 6% (final concentration) trichloroacetic acid (TCA). The pH 4·6 and 6% TCA 13 soluble extracts of milk showed high correlations (R2 > 0·93) by the TNBS, fluorescamine and 14 RP-HPLC methods, confirming increased proteolysis during storage. For gel electrophoresis,15 extensive proteolysis was confirmed by the disappearance of α- and β-casein bands on the seventh 16 day, which was more evident in the highest plasmin concentration. This was accompanied by the 17 appearance of α- and β-casein proteolysis products with higher intensities than on previous days, 18 implying that more products had been formed as a result of casein breakdown. The fluorescamine 19 method had a lower detection limit compared with the other methods, whereas gel electrophoresis 20 was the best qualitative method for monitoring β-casein proteolysis products. Although HPLC was the 21 most sensitive, the TNBS method is recommended for use in routine laboratory analysis on the basis 22 of its accuracy, reliability and simplicity.

Transgenic soya beans: economic implications for EU livestock sector

World oilseed trade consists of many closely substitutable commodities, with canola and cottonseed as possible alternatives to soya beans for many purposes. Transgenic events in all three crops have been widely adopted, particularly in North and South America, for compelling economic or agronomic reasons. Despite the close attention from organizations concerned about the potential consequences of transgenic crop adoption, there appears to be no substantiated evidence of transgenic DNA in meat or milk products when such crops are fed to livestock. The global area of these transgenic crops continues to increase. No transgenic canola, cotton or soya bean crops are permitted for commercial cultivation in Europe, and although transgenic feed resources are permitted for import, importers risk shipments being denied entry if the traces of an unauthorized transgenic crop are detected. These tight controls can mean that livestock farmers in the EU are disadvantaged due to restricted access to cheaper feed or higher feed costs, and they are thus loosing a degree of competitive advantage. This paper reviews the extent to which transgenic soya beans have become the ‘conventional’ method of cultivation elsewhere, and notes implications this has for livestock nutrition, traceability and economics within the EU. The paper concludes with discussion regarding the implications for the EU of delayed acceptance of newly available transgenic traits.