A new sterilization and inoculation method in silage research

The study aimed at evaluating an effective sterilization–inoculation technique to facilitate silage research on the effect of forage microflora on fermentation variables. The sterilization effect of heating at 60°C for 3 h + 103°C for 15 h was tested on samples of grass, grass–clover, white clover and maize, pre‐dried at 60°C to a dry‐matter (DM) content >900 g kg^?1. The ensilability of treated samples, reconstituted to original DM concentration (250–390 g kg^?1), was assessed by inoculation with microfloras extracted from the original samples. Microfloral inoculants were obtained by a combination of centrifugation (15 500  g for 40 min) and filtration (0·45 and 0·22 μm pore sizes) of the supernatant. The sterilization treatment effectively sterilized the forage samples but decreased water soluble carbohydrates by 49% and N buffer solubility by 22% and increased the acid detergent insoluble N proportion of total N by 53% (P < 0·05). The reconstituted silages had 18% less lactic acid, 20% less ethanol and 37% less ammonia‐N (P < 0·05), but volatile fatty acids and 2,3‐butanediol did not differ from the untreated silages (P > 0·05). Counts of lactic acid bacteria, enterobacteria, clostridia, yeasts and moulds in the two silage treatments were also similar (P > 0·05). It is concluded that, despite causing chemical and physical alterations, the sterilization–inoculation technique evaluated could be a useful tool for future studies on the effects of microflora on ensiling results.