Periodic Report Summary - SMETHANE (Technological platform to develop nutritional additives to reduce methane emissions from ruminants)
Project context and objectives:
SMETHANE project aims to provide a technological platform for small and medium sized enterprises (SMEs) to develop and progress further knowledge on the successful use of nutritional additives to reduce methane (CH4) emissions from ruminants. CH4 is the second most important greenhouse gas, after carbon dioxide (CO2) and accounts for nearly one fifth of the global greenhouse effect. Agriculture, waste and energy account for practically all of the European Union's (EU) CH4 emissions, 45 %, 32 % and 23 % respectively.
Reduction options for these sectors have been identified and include a number of promising areas where EU action to reduce CH4 emissions could soon produce significant results. At a global scale, livestock farming contributes up to 18 % of total greenhouse gas emissions. In Europe, almost all livestock-related CH4 emissions arise from fermentation in the digestive tract of herbivorous animals (namely 70 %). The inhibition of CH4 formation by ruminants has long been an objective of ruminant nutritionists, being the development of new feed additives (mainly based on plant extracts) one of the most promising. Despite optimistic announcements in the press, plans for their application appear premature because trials are being performed under conditions that are far removed from practice; mainly in vitro, with a limited number of diets that cannot be applied to the wide range of ruminants production systems in Europe (dairy cattle, beef, sheep and goats raised under both extensive and intensive conditions). The variability and contradictory nature of the results of the results being reported is stopping/restricting the uptake and use of these new compounds in the animal feeding market. The numbers of companies (mainly SMEs) working on developing plants compounds able to reduce CH4 production in the rumen and therefore increase the efficiency of the animal production have significantly increased in the last decade; however, the implementation of these 'developmental' compounds in practical conditions is not yet feasible and no commercial products are currently marketed. The United Nations (UN, see http://unfccc.int/resource/docs/2008/tp/08.pdf online) have identified five main barriers to the development of novel dietary additives to decrease CH4 emissions namely:
1. a lack of suitable laboratory and animal facilities available for testing and development
2. a lack of information on whether the reduction of emissions can be sustained over a long period of time
3. a lack of cost effective delivery mechanisms
4. a lack of knowledge on the wider effects of such additives on the animal and the environment
5. the cost and complexity of generating data to satisfy both national and international regulations.
These problems are particularly pressing for SMEs. Typically SMEs will have only a limited access to research and development facilities, whilst the large scale animal testing facilities needed to measure emissions in animal models are beyond their limited development budgets. Even when SMEs are able to partner with local Universities or Research Institutes this does not provide access to the range of production systems and animal types that would be required for successful product development and registration at a whole EU level.
This proposal is designed to remove the restriction that SMEs face in successfully developing and marketing novel compounds, in particular plant extracts, capable of decreasing CH4 production from ruminant animals. Specifically we will establish research and development platforms that will allow SMEs to determine:
1. the means of delivery of such the compounds in the diet and the stability over time under different storage conditions of such compounds
2. detailed knowledge of the dose response curve for such compounds under different production systems and with different basal diets
3. data on the acceptability of such compounds in target animals and the variability and extent of the mitigation potential in animal trials were CH4 emissions are measured
4. the persistence of the inhibitory effect of such compounds on CH4 production over long periods of time, and
5. the potential 'side effects' such as change in flavour/taste of the final animal product (milk/meat).
Project results:
Overall, the number of compounds tested in the different stages of the SMETHANE technological platform is 13 for stability, 25 in the in vitro screening, 21 for short term, 22 for long term in vitro trials and 5 under on farm conditions.
SMETHANE has addressed the limitation of unknown stability of active molecules in plant extracts by using state of the art analytical tools, such as gas liquid or liquid chromatography combined with mass spectrometry (GC-MS, LC-MS) in work package two (WP2). In WP2 15 different plant extracts have been used to develop standard operational protocols (SOPs) by INRA (France) and CSIC (Spain) for the detection and quantification of the active molecules in a compound feed matrix. The results obtained show a high degree of variation in the recovery of the molecules depending on chemistry and physical presentation of the plant extract. The tested extracts have been incorporated into a feed meal and pelleted diet at around 500 ppm in order to study the stability under three different temperatures (4, 15 and 30 °C) over two months period. As observed for pelleting, loss of molecules was variable depending on the additive with losses increasing at higher temperatures. Residual activity in some compounds was seriously reduced after one month storage at 30 °C. Pelleting process even at low temperature affects negatively the products tested. Concentrate feeds containing plant-derived additives may have a short shelf life under storage conditions found in tropical and subtropical areas or during the hot season in temperate areas.
We have established in vitro and in vivo models to measure effects on CH4 production (WP3 and WP4). In vitro incubations are short term (24h) involving the co-incubation of rumen fluid mixed in an anaerobic buffer with a test diet plus or minus the plant extract at 39 °C under anaerobic conditions. At the end of the incubation gas production is measured and CH4 concentrations determined. SME partners were offered screening of each compound at four concentrations (including controls), two levels of pH and four different basal diets resulting in 32 treatments per compound. Some SME partners opted to screen either a larger number of compounds or a larger range of concentrations and a single diet as an initial screen in order to target more detailed results from subsequent screens. Five compounds have CH4 reduction potential in excess of 40 % but at some concentrations of these also have a detrimental effect of reducing total volatile fatty acids (VFA) production which may affect animal performance due to VFAs providing approximately 85 % of the energy requirements of ruminants. Correlation matrices between compound concentrations and CH4 reduction and VFA production have been produced and results used to design in vivo trials.
In vivo measurements have been made either in calorimeter chambers in which the animal is restrained in a chamber through which air is drawn at a known and accurately determined rate or by placing a bolus that releases sulfur hexafluoride at a known rate into the rumen and by collecting a representative sample of the exhaled air over much of the day, from the sulphur hexafluoride (SF6) release rate plus the SF6/CH4. The results show that high doses applied as in in vitro trials may deplete feed intake, while intermediate (around 100 ppm rumen volume basis) could represent a good compromise. The lack of effect or strong effect of some compounds in short term has been investigated in longer trials over six weeks' periods in cattle (dairy or beef). The results from the long term trials showed that in general the doses that are effective in small ruminants do not show effect in large ruminants and that some blends of essential oils do not work in short term trials (seven days) but become effective after four weeks of treatment, which highlight the importance of performing trials in the medium term.
In WP5 three on farm trials have been conducted to investigate the effect of additives on productivity and product quality: two blends of essential oils and one organosulphour compound. The results on the response from animals treated with essential oils are rather positive (increase in milk production and daily weight gain), while those treated with organosulphur compounds show a numerical increase in weight gain that did not reach statistical significance due to the large variation among groups and the limited number of animals used.
Within WP7 the project web site has been set up (see http://www.smethane.eu online) which contains the most relevant information of the project plus the documents generated over the course of it. As planned in the project proposal three workshops have been ran in there different countries, i.e. on 21 and 22 September 2011 in Birmingham, United Kingdom (UK); on 23 and 24 March 2012, Madrid, Spain; and on 4 and 5 October 2012, Clermont-Ferrand, France. These workshops offered each SME partner the opportunity to invite four customers to follow the progress of the project. The material generated in each workshop is available at the project website (refer to http://www.smethane.eu for details).
Potential impact:
The project is addressing the six major constraints that currently limit the sector of nutritional additives for animal production, specifically:
1. the need to standardise and report the concentration of active component (WP2)
2. stability of the compounds in practical conditions (WP2, WP3)
3. persistence of the effects/adaptation of the rumen ecosystem (WP4)
4. lack of in vivo data over a range of livestock production systems (WP5)
5. effect of extract on the perceived quality of milk products (WP6)
6. a lack of production data on which to base calculations of market prices (WP4, WP5 and WP6).
The results obtained in project have assisted SMEs in the advance knowledge or technological progress by:
1. Describing the effects of pelleting and storage conditions of animals feeds on the stability of the active compounds developed by each SME. The results obtained in this regard show that pelleting process even at low temperature affect negatively the products tested. Also Concentrate feeds containing plant-derived additives may have a short shelf life under storage conditions found in tropical and subtropical areas or during the hot season in temperate areas. The information obtained by each SME will help then to design effective strategies on the application of their compounds in animal feeds in collaboration with feeds sector.
2. For the first time describing the correlation between in vitro and in vivo data (if it is possible to establish such a relationship it will greatly facilitate the development of such products). As stated in previous section the results obtained show no direct correlation between in vitro and in vivo due to a number of factors, as described in deliverable 4.2. This confirms the need to perform enough animal trials to securely test a compound.
3. Describing, for the first time, the variation in response to a mitigation strategy over different production systems and diets. This information is required if we are to fully understand the potential of such an approach to decrease greenhouse gas emissions over Europe, it is also vital for the SME partner in registration of their products). The results generated show the importance of performing trials in which the treatment is applied long enough to truly assess the effectiveness of the compound. This is compulsory when a strategy is being developed for future implementation in practical farming. SMETHANE has provided results on the effectiveness of compounds when they are evaluated in dairy and beef cattle systems, both very important in Europe. In addition, the small ruminants sector will benefit from results obtained using sheep and goats. This is very important when the use of a specific compound is considered within the farming sector across the continent.
4. Addressing concerns over transfer of residues into milk (again vital both in terms of public acceptability and registration). Two different types of compounds have been tested for their transfer into milk in dairy cows, namely blend essential oils and allicin. Under the conditions of the trials performed in SMETHANE, no transfer of the active molecules in the blend of essential oils or their metabolites is detected in milk of dairy cows, while clearly organosulhphur compounds taint milk. These results have obvious social implications for both the interest of the company to further develop their application and for the consumer.
5. An online calculator developed within WP6 has been provided as a demonstration tool in the project website, although it does not contain values associated with a particular SME partners' product. It is intended to help assist in marketing and sales of the SME partners who have additional data to support their findings. All assumptions to our knowledge are sound and are acceptable at time of production. The calculator can be modified by individual SME partners in collaboration with the research and technological development (RTD) authors. The calculator provides an indication based upon the information supplied. It does not perform a life cycle assessment but can be used to generate data more scientifically than a tier one approach.
Project website: http://www.smethane.eu
SMETHANE project aims to provide a technological platform for small and medium sized enterprises (SMEs) to develop and progress further knowledge on the successful use of nutritional additives to reduce methane (CH4) emissions from ruminants. CH4 is the second most important greenhouse gas, after carbon dioxide (CO2) and accounts for nearly one fifth of the global greenhouse effect. Agriculture, waste and energy account for practically all of the European Union's (EU) CH4 emissions, 45 %, 32 % and 23 % respectively.
Reduction options for these sectors have been identified and include a number of promising areas where EU action to reduce CH4 emissions could soon produce significant results. At a global scale, livestock farming contributes up to 18 % of total greenhouse gas emissions. In Europe, almost all livestock-related CH4 emissions arise from fermentation in the digestive tract of herbivorous animals (namely 70 %). The inhibition of CH4 formation by ruminants has long been an objective of ruminant nutritionists, being the development of new feed additives (mainly based on plant extracts) one of the most promising. Despite optimistic announcements in the press, plans for their application appear premature because trials are being performed under conditions that are far removed from practice; mainly in vitro, with a limited number of diets that cannot be applied to the wide range of ruminants production systems in Europe (dairy cattle, beef, sheep and goats raised under both extensive and intensive conditions). The variability and contradictory nature of the results of the results being reported is stopping/restricting the uptake and use of these new compounds in the animal feeding market. The numbers of companies (mainly SMEs) working on developing plants compounds able to reduce CH4 production in the rumen and therefore increase the efficiency of the animal production have significantly increased in the last decade; however, the implementation of these 'developmental' compounds in practical conditions is not yet feasible and no commercial products are currently marketed. The United Nations (UN, see http://unfccc.int/resource/docs/2008/tp/08.pdf online) have identified five main barriers to the development of novel dietary additives to decrease CH4 emissions namely:
1. a lack of suitable laboratory and animal facilities available for testing and development
2. a lack of information on whether the reduction of emissions can be sustained over a long period of time
3. a lack of cost effective delivery mechanisms
4. a lack of knowledge on the wider effects of such additives on the animal and the environment
5. the cost and complexity of generating data to satisfy both national and international regulations.
These problems are particularly pressing for SMEs. Typically SMEs will have only a limited access to research and development facilities, whilst the large scale animal testing facilities needed to measure emissions in animal models are beyond their limited development budgets. Even when SMEs are able to partner with local Universities or Research Institutes this does not provide access to the range of production systems and animal types that would be required for successful product development and registration at a whole EU level.
This proposal is designed to remove the restriction that SMEs face in successfully developing and marketing novel compounds, in particular plant extracts, capable of decreasing CH4 production from ruminant animals. Specifically we will establish research and development platforms that will allow SMEs to determine:
1. the means of delivery of such the compounds in the diet and the stability over time under different storage conditions of such compounds
2. detailed knowledge of the dose response curve for such compounds under different production systems and with different basal diets
3. data on the acceptability of such compounds in target animals and the variability and extent of the mitigation potential in animal trials were CH4 emissions are measured
4. the persistence of the inhibitory effect of such compounds on CH4 production over long periods of time, and
5. the potential 'side effects' such as change in flavour/taste of the final animal product (milk/meat).
Project results:
Overall, the number of compounds tested in the different stages of the SMETHANE technological platform is 13 for stability, 25 in the in vitro screening, 21 for short term, 22 for long term in vitro trials and 5 under on farm conditions.
SMETHANE has addressed the limitation of unknown stability of active molecules in plant extracts by using state of the art analytical tools, such as gas liquid or liquid chromatography combined with mass spectrometry (GC-MS, LC-MS) in work package two (WP2). In WP2 15 different plant extracts have been used to develop standard operational protocols (SOPs) by INRA (France) and CSIC (Spain) for the detection and quantification of the active molecules in a compound feed matrix. The results obtained show a high degree of variation in the recovery of the molecules depending on chemistry and physical presentation of the plant extract. The tested extracts have been incorporated into a feed meal and pelleted diet at around 500 ppm in order to study the stability under three different temperatures (4, 15 and 30 °C) over two months period. As observed for pelleting, loss of molecules was variable depending on the additive with losses increasing at higher temperatures. Residual activity in some compounds was seriously reduced after one month storage at 30 °C. Pelleting process even at low temperature affects negatively the products tested. Concentrate feeds containing plant-derived additives may have a short shelf life under storage conditions found in tropical and subtropical areas or during the hot season in temperate areas.
We have established in vitro and in vivo models to measure effects on CH4 production (WP3 and WP4). In vitro incubations are short term (24h) involving the co-incubation of rumen fluid mixed in an anaerobic buffer with a test diet plus or minus the plant extract at 39 °C under anaerobic conditions. At the end of the incubation gas production is measured and CH4 concentrations determined. SME partners were offered screening of each compound at four concentrations (including controls), two levels of pH and four different basal diets resulting in 32 treatments per compound. Some SME partners opted to screen either a larger number of compounds or a larger range of concentrations and a single diet as an initial screen in order to target more detailed results from subsequent screens. Five compounds have CH4 reduction potential in excess of 40 % but at some concentrations of these also have a detrimental effect of reducing total volatile fatty acids (VFA) production which may affect animal performance due to VFAs providing approximately 85 % of the energy requirements of ruminants. Correlation matrices between compound concentrations and CH4 reduction and VFA production have been produced and results used to design in vivo trials.
In vivo measurements have been made either in calorimeter chambers in which the animal is restrained in a chamber through which air is drawn at a known and accurately determined rate or by placing a bolus that releases sulfur hexafluoride at a known rate into the rumen and by collecting a representative sample of the exhaled air over much of the day, from the sulphur hexafluoride (SF6) release rate plus the SF6/CH4. The results show that high doses applied as in in vitro trials may deplete feed intake, while intermediate (around 100 ppm rumen volume basis) could represent a good compromise. The lack of effect or strong effect of some compounds in short term has been investigated in longer trials over six weeks' periods in cattle (dairy or beef). The results from the long term trials showed that in general the doses that are effective in small ruminants do not show effect in large ruminants and that some blends of essential oils do not work in short term trials (seven days) but become effective after four weeks of treatment, which highlight the importance of performing trials in the medium term.
In WP5 three on farm trials have been conducted to investigate the effect of additives on productivity and product quality: two blends of essential oils and one organosulphour compound. The results on the response from animals treated with essential oils are rather positive (increase in milk production and daily weight gain), while those treated with organosulphur compounds show a numerical increase in weight gain that did not reach statistical significance due to the large variation among groups and the limited number of animals used.
Within WP7 the project web site has been set up (see http://www.smethane.eu online) which contains the most relevant information of the project plus the documents generated over the course of it. As planned in the project proposal three workshops have been ran in there different countries, i.e. on 21 and 22 September 2011 in Birmingham, United Kingdom (UK); on 23 and 24 March 2012, Madrid, Spain; and on 4 and 5 October 2012, Clermont-Ferrand, France. These workshops offered each SME partner the opportunity to invite four customers to follow the progress of the project. The material generated in each workshop is available at the project website (refer to http://www.smethane.eu for details).
Potential impact:
The project is addressing the six major constraints that currently limit the sector of nutritional additives for animal production, specifically:
1. the need to standardise and report the concentration of active component (WP2)
2. stability of the compounds in practical conditions (WP2, WP3)
3. persistence of the effects/adaptation of the rumen ecosystem (WP4)
4. lack of in vivo data over a range of livestock production systems (WP5)
5. effect of extract on the perceived quality of milk products (WP6)
6. a lack of production data on which to base calculations of market prices (WP4, WP5 and WP6).
The results obtained in project have assisted SMEs in the advance knowledge or technological progress by:
1. Describing the effects of pelleting and storage conditions of animals feeds on the stability of the active compounds developed by each SME. The results obtained in this regard show that pelleting process even at low temperature affect negatively the products tested. Also Concentrate feeds containing plant-derived additives may have a short shelf life under storage conditions found in tropical and subtropical areas or during the hot season in temperate areas. The information obtained by each SME will help then to design effective strategies on the application of their compounds in animal feeds in collaboration with feeds sector.
2. For the first time describing the correlation between in vitro and in vivo data (if it is possible to establish such a relationship it will greatly facilitate the development of such products). As stated in previous section the results obtained show no direct correlation between in vitro and in vivo due to a number of factors, as described in deliverable 4.2. This confirms the need to perform enough animal trials to securely test a compound.
3. Describing, for the first time, the variation in response to a mitigation strategy over different production systems and diets. This information is required if we are to fully understand the potential of such an approach to decrease greenhouse gas emissions over Europe, it is also vital for the SME partner in registration of their products). The results generated show the importance of performing trials in which the treatment is applied long enough to truly assess the effectiveness of the compound. This is compulsory when a strategy is being developed for future implementation in practical farming. SMETHANE has provided results on the effectiveness of compounds when they are evaluated in dairy and beef cattle systems, both very important in Europe. In addition, the small ruminants sector will benefit from results obtained using sheep and goats. This is very important when the use of a specific compound is considered within the farming sector across the continent.
4. Addressing concerns over transfer of residues into milk (again vital both in terms of public acceptability and registration). Two different types of compounds have been tested for their transfer into milk in dairy cows, namely blend essential oils and allicin. Under the conditions of the trials performed in SMETHANE, no transfer of the active molecules in the blend of essential oils or their metabolites is detected in milk of dairy cows, while clearly organosulhphur compounds taint milk. These results have obvious social implications for both the interest of the company to further develop their application and for the consumer.
5. An online calculator developed within WP6 has been provided as a demonstration tool in the project website, although it does not contain values associated with a particular SME partners' product. It is intended to help assist in marketing and sales of the SME partners who have additional data to support their findings. All assumptions to our knowledge are sound and are acceptable at time of production. The calculator can be modified by individual SME partners in collaboration with the research and technological development (RTD) authors. The calculator provides an indication based upon the information supplied. It does not perform a life cycle assessment but can be used to generate data more scientifically than a tier one approach.
Project website: http://www.smethane.eu