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In vitro methods have been standardized and tested to correctly simulate the rumen environment and fermentation process. A few studies have verified that the feed degradability achieved as a result of stirring the samples is higher when the samples are incubated under continuous stirring than when they are only stirred twice daily. The objective of this study has been to verify the effect of the speed of stirring on feed degradability during In vitro incubation. For this purpose, the apparent and true dry matter degradability (ADMD and TDMD) of grass hay, pelleted alfalfa, corn silage, barley meal, straw, and a total mixed ration (TMR) were measured after 48 h of incubation in jars under different rotation speeds. The same types of feed were placed in the four jars of each instrument, and the rotation system of the machine was modified to ensure the simultaneous rotation of a pair of original jars (which sometimes stopped and/or rotated slowly and irregularly) together with a pair of modified jars under regular and continuous rotation. A rev counter data logger was mounted onto the jars, and the rotation speeds of the original and modified jars were measured and compared under different conditions (empty jars, jars with liquid, jars with rumen fluid, and sample bags). The modifications to the instruments stabilized the rotation of the jars, thereby making the stirring more regular during incubation. The degradability was partly influenced by the regular stirring, albeit with just one instrument, and for grass hay, barley meal, corn silage, and TMR. In short, it has been found that the regular stirring of sample bags is not essential to obtain reliable degradability measurement during incubation, although it is better to maintain a constant rotation to ensure a regular and standardized In vitro incubation process and therefore to allow reproducibility and comparisons of the results on feed degradability.

This review summarises the use of the Ankom DaisyII incubator (ADII; Ankom Technology Corporation Fairport, NY, USA), as presented in studies on digestibility, and its extension to other species apart from ruminants, from its introduction until today. This technique has been modified and adapted to allow for different types of investigations to be conducted. Researchers have studied and tested different procedures, and the main sources of variation have been found to be: the inoculum source, sample size, sample preparation, and bag type. In vitro digestibility methods, applied to the ADII incubator, have been reviewed, the precision and accuracy of the method using the ADII incubator have been dealt with, and comparisons with other methods have been made. Moreover, some hypotheses on the possible evolutions of this technology in non-ruminants, including pets, have been described. To date, there are no standardised protocols for the collection, storage, and transportation of rumen fluid or faeces. There is also still a need to standardise the procedures for washing the bags after digestion. Moreover, some performance metrics of the instrument (such as the reliability of the rotation mechanism of the jars) still require improvement.

Apparent in vitro dry matter digestibilities of selected ruminant feeds were determined with the DAISYII incubator (ADD, ANKOM Technology Corp., Macedon, NY) and the Tilley and Terry technique (ADTT). True in vitro dry matter digestibility was also determined with the DAISYII incubator (TDD). The ADD and ADTT did not differ for grain crop silages and total mixed rations. The ADD was 9.0 percentage points higher than ADTT for grains and 3.4 percentage points lower than ADTT for grass and legume forages. The TDD was between 5.7 and 11.2 percentage points higher than ADD depending on the feed. Key words: In vitro dry matter digestibility, forages, grains, ANKOM DAISYII incubator