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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.

The apparent dry matter digestibility of diets for rabbits was measured in vivo (ADMDvv) and in vitro with the Ankom DaisyII Incubator. Four diets were tested: low fiber (LF), LF + 5% of pregerminated fenugreek seeds (PGFS) (LF5), LF + 10% PGFS (LF10), and high fiber (HF). For the ADMDvv, feces samples were collected from 56 White New-Zealand × Californian rabbits fed the 4 diets, animals were randomly allocated into 4 groups and housed in individual cages. For the in vitro trial, 3 methods were tested: fecal inoculum (FA) with Kansans State buffer, fecal inoculum with artificial saliva (FB), and multienzyme (ENZ). Fecal inocula were collected at slaughtering from the distal colon of rabbits fed ad libitum the LF diet. For FA and FB methods, the digestibility was measured at 36 and 48 h. The in vitro methods ranked the apparent dry matter digestibility of diets in the same order as in vivo, but ENZ values were always higher than FA and FB at 36 and 48 h. The prediction equations of in vivo digestibility with the ENZ method showed higher coefficient of determination (R2 = 0.69) and lower SE (0.190) than FA and FB, also, reproducibility was higher with ENZ (CV = 3.1%). In conclusion, different methods can be applied to the Ankom DaisyII Incubator to study the digestibility in rabbits. In our trial, the better reproducibility was observed with the multienzyme method than FA and FB were probably related to the variability of inocula.

A study was conducted to determine the reliability of the novel in vitro dry matter digestibility (IVDMD) method for predicting the in vivo organic matter digestibility (OMD) of forages. The study was carried out on two sets of feeds of known OMD determined on sheep: set A (n = 35), consisting of whole crop cereal (corn and barley) herbages and set B (n = 80), mostly consisting of grass and clover silages. IVDMD was determined by the CEL48-ND method in which 500 mg of dried and ground feed sample was inserted into filter bag (ANKOM Technology Corporation, Fairport, NY, USA) and then the bags were incubated for 48 h at 39 °C in the cellulase solution (celullase Onozuka R10), in the jars of DaisyII Incubator (ANKOM Technology Corporation). After incubation the bags with residues were extracted in a neutral detergent (ND) for 1 h at 100 °C in Ankom220 Fiber Analyzer (ANKOM Technology Corporation). The data for all sets were used to find the best single and multiple regression equations to predict OMD. For each set the linear regressions were calculated in which OMD was the dependent variable (Y) whereas IVDMD determined by CEL48-ND was the predictor X1 and contents of nutrients (g/kg DM) were predictors X2, X3, X4 or X5. The equations were compared by means of coefficient of determination (R2), standard deviation of differences between observed (Yi) and predicted value (Ỹi) (residual standard deviation; RSD), and mean square prediction error (MSPE). Additionally, the verification of equations was also done based on the Akaike information criterion (AIC). Mean R2 for the regression equations of OMD by CEL48-ND was 0.468 (set A), 0.581 (set B) and 0.539 (set A + B). Including the next predictor (X2, X3, X4 or X5) increased R2, and decreased RSD, MSPE and AIC, especially in the equations calculated for whole crop corn herbages, grass silages and clover silages. It can be concluded that the in vitro method presented in this study is a simple alternative for existing methods in which buffered rumen fluid is used. Using a standard enzyme available commercially worldwide may decrease variation between laboratories. Further, using filter bags and Daisy Incubator decreases labour costs and use of animals.AbbreviationsAIC, akaike information criterion; ADF, acid detergent fibre; aNDF, neutral detergent fibre; AFBT, ANKOM filter bag technique; CP, crude protein; CS, subset ‘clover silages’; DK, Denmark; DM, dry matter; FIN, Finland; GS, subset ‘grass silages’; IVDMD, in vitro dry matter digestibility; MSPE, mean square prediction error; ND, neutral detergent solution; OMD, organic matter digestibility; R2, coefficient of determination; RSD, residual standard deviation; WCB, subset ‘whole crop barley’; WCC, subset ‘whole crop corn’

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