Low Emission Feed : using feed additives to decrease methane production in dairy cows

Research into manipulating methane (CH4) production as a result of enteric fermentation in ruminants currently receives global interest. Using feed additives may be a feasible strategy to mitigate CH4 as they are supplied in such amounts that the basal diet composition will not be largely affected. The latter is relevant because ruminants have the capacity to convert human inedible feedstuffs into human edible energy and protein. However, the application of CH4 mitigation feed additives may be hampered by several negative side effects including trade-offs with other environmental impacts, negative effects on animal performance, and lack of persistency of the mitigating effect. The research described in this thesis addresses both the mitigating effect of feed additives as well as its persistency. The main focus was on investigating additivity of the CH4 mitigating effect of feed additives, on the adaptation of rumen microbes to long term feeding of feed additives, and on exploring the potential of rotational feeding of additives to avoid (or reduce) microbial adaptation. In an experiment with lactating dairy cows in climate respiration chambers to study potential interactions between the effects of feeding nitrate and docosahexaenoic acid (DHA; C22:6 n-3) on enteric CH4 production, the effects of nitrate and DHA on CH4 yield [g/kg dry matter intake (DMI)] and CH4 intensity [g/kg fat- and protein- corrected milk (FPCM)], were additive (<strong>Chapter 2</strong>). Nitrate decreased CH4 irrespective of the unit in which it was expressed, and the average decline in CH4 emission corresponds to 85% of the stoichiometric potential of nitrate to decrease CH4. Feeding DHA had no effect on CH4 yield, but resulted in a higher CH4 intensity, because of milk fat depression. The interaction effect between nitrate and DHA on fiber digestibility indicated that negative effects of nitrate on apparent total tract digestibility of nutrients were alleviated by DHA, probably due to an altered feed intake pattern. Using an isotope measurement protocol in the same study, it was demonstrated that effects of nitrate as a CH4 mitigating feed additive on fiber degradation in the rumen can be detected by evaluating diurnal patterns of 13C enrichment of exhaled CO2 (<strong>Chapter 3</strong>). Feeding nitrate, but not DHA, resulted in a pronounced increase in 13C enrichment of CO2 in the first 3 to 4 h after feeding only. Results support the hypothesis that effects of a feed additive on the rate of fiber degradation in the rumen can be detected by evaluating diurnal patterns of 13C enrichment of CO2. A prerequisite for this detection method is that the main ration components differ in natural 13C enrichment (e.g., C3 and C4 plants), and in content of the nutrients that are expected to be involved in a shift in fermentation (e.g., starch and fiber) or in degradability of a nutrient. In a combined in vivo and in vitro trial, the adaptation to CH4 mitigating feed additives, viz. an essential oil blend or lauric acid (C12:0), compared with a control diet was first investigated using the in vitro gas production technique during the period that lactating cows were adapting to certain feed additives (<strong>Chapter 4</strong>). Rumen fluid was collected from each cow at several days relative to the introduction of the additives in the diets and used as inoculum for the gas production experiment with each of the three different substrates that reflected the treatment diets offered to the cows. The feed additives in the donor cow diet had a stronger effect on in vitro gas and CH4 production than the same additives in the incubation substrate. From day 4 onwards, the C12:0 diet persistently reduced gas and CH4 production, total volatile fatty acid concentration, acetate molar proportion and in vitro organic matter degradation, and increased propionate molar proportion. In contrast, in vitro CH4 production was reduced by the essential oils diet on day 8, but not on days 15 and 22. In line with these findings, the molar proportion of propionate in fermentation fluid was higher, and that of acetate smaller, for the essential oils diet than for the control diet on day 8, but not on days 15 and 22. Overall, the data indicate a transient effect of the essential oils on CH4 production, which may indicate microbial adaptation, whereas the CH4 mitigating effect of C12:0 persisted. It is recommended that this phenomenon is considered in the planning of future studies on the mitigation potential of feed additives in vitro. In a follow-up in vivo study, it was investigated whether the alternate feeding of two CH4 mitigating feed additives with a different mode of action (viz. C12:0 and a blend of essential oils) would result in a persistently lower CH4 production compared to feeding a single additive over a period of 10 weeks. The experiment comprised a pre-treatment period and three two-week measurement periods, with two periods of 2 weeks in between in which CH4 emission was not measured. Cows received either continuously the essential oil blend, or both the essential oil blend and C12:0 following a weekly rotation schedule (<strong>Chapter 5</strong>). Both CH4 yield and CH4 intensity changed over time, but were not affected by treatment. Methane yield and intensity were significantly lower (12 and 11%, respectively) in period 1 compared with the pre-treatment period, but no significant difference relative to the pre-treatment period was observed in period 3 (numerically 9 and 7% lower, respectively) and in period 5 (numerically 8 and 4% lower, respectively). These results indicate a transient decrease in CH4 yield and intensity in time, but no improvement in extent or persistency of CH4 reduction due to rotational feeding of essential oils and C12:0 in lactating dairy cows. However, there were indications that the concept of rotation may be effective and warrants further investigation. The additives and concepts tested in this thesis are applied under specific experimental conditions. More mechanistic understanding is required to predict the response of the same additives when supplemented to other basal diets or cows in a different physiological state. Trade-offs in environmental impact, and effects on cow health and performance, and on milk processing parameters and food safety are important aspects to consider in future research on the application of feed additives as CH4 mitigation strategy.