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Abstract
In vitro fermentation is a useful method to test the effects of secondary metabolites on the rumen fermentation. It can be a noninvasive method when the rumen fluid is collected from slaughterhouses from dead animals. The experimental design for this type of experiment is normally a factorial design where common factors tested are: incubation time, main energy source incubated, and additive concentrations (secondary metabolites). The data gathered from the incubations can be quite extensive and therefore the statistical power of the in vitro method can be very good. In this paper we will discuss the main characteristics (how the method works and which parameters are measured) of the in vitro fermentation method.
Highlights
Noninvasive methods to study novel feedstuff to be used in animal agriculture have currently been developed and improved
VFA and adenosine triphosphate (ATP) are used as the available energy sources for the animal, while microbial cells are the significant source of quality protein entering the small intestine (Demeyer, 1981; Russel & Hespell, 1981)
The incubation method works as follows: about 1 g of ground energy source sample, plus the secondary metabolite under test in different inclusion levels according to experimental design, are put into 50-ml polypropylene tubes and 40 ml of the buffered inoculum (32 ml of buffer solution plus 8 ml of filtered rumen fluid) dispensed into each tube, purged with CO2 to maintain anaerobic conditions, sealed with rubber stoppers fitted with gas pressure release valves, and incubated in a temperature controlled shaking water bath (39 oC) (Figure 1)
Summary
Noninvasive methods to study novel feedstuff to be used in animal agriculture have currently been developed and improved. The incubation will yield vast datasets with information such as: pH, ammonia levels, methane concentration, volatile fatty acids, dry matter degradability, total gas production, profile of microbial population, amongst others (Calsamiglia et al, 2007; Makkar et al, 2006; Yang et al, 2010). The analysis of this information can reveal the effects of an individual chemical compound when incubated in the rumen
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