BIOREFINERY GLAS

D3.4 Pre-commercial product

specifications and regulatory
compliance action plans

BIOREFINERY GLAS IS A EUROPEAN INNOVATION PARTNERSHIP (EIP) FUNDED BY THE DEPARTMENT OF AGRICULTURE, FOOD,
AND THE MARINE (DAFM) UNDER THE RURAL DEVELOPMENT PROGRAMME 2014-2020
 Deliverable 3.4
Pre-commercial product specifications
and regulatory compliance action plans
Project title: Biorefinery Glas
Contract Nº: LLOC2043
Start date: 01/02/2019
Duration: 24 months

Deliverable number D 3.4

Deliverable title Pre-commercial product specifications and
regulatory compliance action plans
Submission due date Date deliverable is due 30/11/2020
Actual Completion date 31/12/2020
Work Package WP 3
WP Lead Beneficiary UCD
Dissemination Level Public
Version 1
Authors Abhay Menon (MTU), James Gaffey (MTU),
Helena McMahon (MTU)
 DOCUMENT CONTROL P AGE

Author(s) First Name/last name (Partner Acronym)

# Reviewer Comments
1 Consortium n/a
Version
history

Comments

Draft
Status
x FINAL
 1. Executive Summary
This report summarizes the technical specifications and regulatory compliance
action plans to be followed for each value-added product generated from
Biorefinery Glas project. In an Irish context, pre-commercialization regulatory
protocols meant for food/feed/nutraceutical applications are reliant on the norms
set aside by the European union. Several regulatory verticals under the
Department of Agriculture, Food and the Marine overlooks the regulatory
compliance action plans required within the Irish market. Products generated from
the project are compared with commercially available market standards, in turn
substantiating the requirement to pitch these products in the market to promote
organic/non-GMO alternatives for Ireland to be in line with EU Biodiversity Strategy
for 2030 1.
Figure 1 illustrates the array of products from Biorefinery Glas- which has immense
potential to be commercialized. The small scale biorefinery unit separates the grass
into two main components – grass juice (liquid fraction) & press cake (solid
fraction). The biorefinery process converts freshly harvested grass supplied by local
farmers into a range of products, including an optimized cattle feed fibre, a non-
GMO protein concentrate feed for monogastrics, a high-value sugar stream of
fructo-oligosaccharides and a grass whey for fertilizer or bioenergy applications.
The project targets a 40% increase in usable protein per hectare and expects to
achieve a 25% reduction in nitrogen emissions in cattle excrement, with additional
emissions savings through displacement of soybean feed imports with a grass-
based monogastric feed. The project also demonstrates and evaluates an
innovative business model for farm diversification into the circular economy and
supports farmers with a range of knowledge exchange and dissemination. From
the juice fraction after refining stage, the proteins are separated from it, the
remaining whey are mainly used for extraction Fructo-oligosaccharides (FOS), and
retentates from the FOS extraction process are pitched in as substrates with
potential for use as, feedstock for anaerobic digestion and in production
biofertilizer.

1
https://eur-lex.europa.eu/legalcontent/EN/TXT/?qid=1590574123338&uri=CELEX:52020DC0380
Figure 1: Products obtained from Biorefinery Glas-small scale refining process
Contents
1. Executive Summary................................................................................................ 2
2. Acronyms and Abbreviations ................................................................................. 5
2.1. Fructo-oligosaccharides ...................................................................................... 6
2.1.1. Introduction ................................................................................................. 6
2.1.2. Technical specifications and comparison with commercial FOS products ..... 7
2.1.3. Regulatory compliance measures ................................................................. 9
2.2. Protein feed for Monogastric animals ............................................................... 12
2.2.1. Introduction ............................................................................................... 12
2.2.2. Technical specification and product comparison with SBM ........................ 12
2.2.3. Regulatory compliance measures ............................................................... 14
2.3. Press cake as an alternative for grass silage ...................................................... 16
2.3.1. Introduction ............................................................................................... 16
2.3.2. Technical specifications and performance of press cake against grass silage
in cattle milk production ...................................................................................... 16
2.3.3. Regulatory compliance measures……………………………………………………………...17

2.4. Biofertilizer production & Digestates in Anerobic digestion .............................. 19

2.4.1. Introduction ............................................................................................... 19
2.4.2. Technical specifications of grass whey fertilizer. ......................................... 19
2.4.3. Regulatory compliance measures ............................................................... 21
3. Conclusions.......................................................................................................... 22
2. Acronyms and Abbreviations
D3.4 Deliverable 3.4
DAFM Department of Agriculture, Food and the Marine
WP Work Package
GHG Green House Gas
N Nitrogen
P Phosphorus
K Potassium
LESS Low Emission Slurry Spreading
DM Dry Matter
FOS Fructooligosaccharides
2.1. Fructo-oligosaccharides

2.1.1. Introduction
Nowadays consumers are becoming more health conscious and the demand for
functional foods that can provide some additional health benefits. Therefore, food
products are increasingly supplemented with prebiotics, resulting in a large commercial
interest in FOS. Fructans are classified as soluble fibres and are available as a food
supplement. Fructans are touted to have the capability of promoting the growth of
beneficial bacteria within the gastrointestinal tract and may provide relief from
2
inflammatory bowel disease . 1-Kestose (isokestose) and nystose are inulin
oligosaccharides comprised of one or two β-1,2-linked fructose residues, respectively,
attached to sucrose. They are obtained from sucrose using a fructosyltransferase and
are marketed as low-calorie, non-cariogenic sweeteners3. Fructan biosynthesis takes
place in the vacuole of the plant cell and uses 1-kestose as a primer. Short-chain FOS
preparations currently used as food ingredients are generally produced by enzymatic
synthesis from sucrose (glucose + fructose; GF) and are characterized as short-chain
mixtures of fructose oligomers of which one (GF2; 1-kestose), two (GF3; nystose), or
three (GF4; β-fructofuranosyInystose) additional fructose units have been added by β2-
1 glycosidic linkages to the fructose unit of sucrose. Figure 2, summarizes the three
different short chain fructo-oligosaccharide sugars obtained from the project

2
J Physiol Biochem. 2009 Sep;65(3):315-28. doi: 10.1007/BF03180584

3
Immunology. 2018. Volume 1: Immunotoxicology, Immunopathology, and Immunotherapy, 203-224.
https://doi.org/10.1016/B978-0-12-809819-6.00014-9

6
 Figure 2: Illustration of short chain FOS units generated from biorefining grass4

The sc-FOS extracts obtained from Biorefinery Glas have the potential to be pitched as
alternative to prebiotic-products with several health claims as projected in the market.
The following sections shall entail the technical specification and prebiotic performance
of these products with commercially available sources.

2.1.2. Technical specifications and comparison with commercial FOS products

The global prebiotics market is expected to reach USD 7.11 billion by 20245. Rising
investment in the food & beverage industry for innovation and development of new
food products enriched with prebiotics coupled with rising demand in dietary
supplements application particularly in infant foods is expected to propel market growth
over the forecast period. In addition, rising use of prebiotics in animal feed is expected
to drive the demand. Growing emphasis on improving overall health is also expected to
fuel product demand over the next eight years. Emergence of prebiotics as an alternate
to antibiotics is expected to fuel demand in animal feed applications. Increasing dairy
production is expected to have a positive impact on market over the next eight years.

On the consumer side, fructo-oligosaccharides are mainly distributed in Europe, China,

Japan, and North America. Among them, Japan is the world’s largest consumer market.

4
Díez-Municio M, de las Rivas B, Jimeno ML, Muñoz R, Moreno FJ, Herrero M. Enzymatic synthesis and characterization of
fructooligosaccharides and novel maltosylfructosides by inulosucrase from Lactobacillus gasseri DSM 20604. Appl Environ Microbiol.
2013 Jul;79(13) 4129-4140. doi:10.1128/aem.00854-13. PMID: 23645191; PMCID: PMC3697562.
5
Fructooligosaccharides (FOS) Market Analysis By Source (Inulin, Sucrose) By Application (Food & Beverages, Infant Formula,
Dietary Supplements, Animal Feed, Pharmaceuticals) And Segment Forecasts To 2024 GVR

7
In 2017, Japan sales accounted for 24.72% of the global market share. The leading
manufacturers are Quantum Hi-Tech, Bowling Bio, and Meiji Japan. The main European
producers are Beneo, Sensus and Japan’s Meiji joint venture BMI in Europe. In Japan,
synthesized FOS using enzymes have been in the market since 1984. The first FOS
industrial producer was the Meiji Seika Kaisha, Ltd. with the product named Meioligo .
FOS produced by Meiji were named “Neosugar” 6. Meioligo products are
commercialized as several product types with different purity level. Currently in Japan,
FOS can be found in more than 500 food products 7. In 2007, the retail FOS market was
estimated to be about US$200/kg. Other companies also commercializing FOS
synthesized using enzymes are GT Nutrition in the United States with NutraFlora®; Cheil
Foods and Chemicals in Korea with OligoSugar; and Victory in China with Beneshine® .
FOS are commercialized with a purity level above 95% (supplier info). Table 1 represents
the list of various market leaders in production of FOS, degrees of polymerization of
each product and their countries of origin.

Table 1: List of market leaders in FOS production and the degrees of polymerization of
each product

Company Name Country Product name Type of Fructan

Tokyo,
Meiji Seika Kaisha Meioligo FOS (DP 3-5)
Japan
Golden
GTC Nutrition Colorado, NutraFlora® FOS (DP < 5)
US
Cheil Foods and Seoul,
Oligo-Sugar FOS (DP 2-4)
Chemicals Korea
Victory Biology Shangai, Beneshine™
FOS (10-20)
Engineering China Ptype
Brussels, Inulin (DP 20-60) and FOS (DP
BENEO-Orafti Orafti®
Belgium 2-8)
Warcoing, Fibruline® Inulin (DP 30-60)
(Cosucra
Belgium Fibrulose® FOS (DP~ 10)
Ayotlan,
Nutriagaves de OLIFRUCTINE- Inulin (DP 3-60) and FOS (DP 3-
Jalisco,
Mexico SP® 5)
Mexico

6
Oku, Tsuneyuki, et al. "Bioavailability and laxative threshold of 1-kestose in human adults." Dynamic Biochem Process
Biotech Mol Bio 3.S1 (2009): 90-5.
7
Macfarlane, G. T., H. Steed, and S. Macfarlane. "Bacterial metabolism and health‐related effects of galacto‐
oligosaccharides and other prebiotics." Journal of applied microbiology 104.2 (2008): 305-344.

8
 Borchwerf
, The
Sensus Frutalose ® FOS (DP 2-25)
Netherlan
ds
Chennai,
Tata Chemicals FOSSENCE® FOS (DP 3-5)
India
Ma'anit,
Galam Ltd GOFOS® FOS (DP avg 4.5)
Israel
Beghin-Meiji Paris, Actilight® FOS (Actilite DP 2-4); (Profeed
Industries France Profeed® DP 3-5)

In Europe, FOS synthesized using enzymes are only commercialized by Beghin-Meiji

Industries. These FOS are produced from sucrose by the company TEREOS sugar group
in France. Beghin-Meiji commercializes FOS for two industry segments: the Actilight® for
food industry, and the Profeed® for animal feed industry. Actilight® is available in
powder and liquid form, with a fibre content ranging from 55% to 95%. Inulin and
oligofructose extracted from the chicory root are produced in Europe by Beneo-Orafti,
Cosucra and Sensus. Cosucra commercializes it by the name of Fibruline® and
Fibrulose®8; Beneo-Orafti as Orafti® ingredients; and Sensus as Frutafit® inulin and
Frutalose®9 oligofructose. By varying the contents in dietary fibre and sugar, these
companies can offer more than one type of product. In Mexico, Nutriagaves de Mexico
S.A. de C.V. (Namex) produces and commercializes inulin and oligofructose extracted
from the desert succulent blue agave (flower) under the brand name OLIFRUCTINE.
Oligofructose is an inulin-type fructan. Oligofructose is derived from Inulin through
partial enzymatic hydrolysis. This conversion is a process that occurs naturally within the
chicory root towards the end of the harvest. Orafti® Oligofructose can therefore be said
to be of 100% vegetable origin. After enzymatic hydrolysis, purification is continued to
meet even the highest quality requirements, including those for infant nutrition.

2.1.3. Regulatory compliance measures

Several regulatory bodies around the world has reviewed and inspected various health
claims projected by manufactures of FOS enriched feed/ food or nutraceutical products.
Figure 3, represents the various regulatory bodies- around the world, which has scoped
and reviewed the claims put forward by various FOS enriched pre-commercial products.

8
https://www.innovafood.net/en/products/cosucra/fibruline-fibrulose
9
https://www.inspiredbyinulin.com/inulin/frutafit-frutalose.html

9
 Figure 3: List of various regulatory bodies inspecting health claims of FOS

This report shall mainly focus on EU regulation namely; European Food Safety Authority
(EFSA; EC) No 178/2002) which proposes on guidelines and communicates on existing
and emerging risks associated with use of any pre-commercial product in food-chain.
EFSA adheres to several principles and practices aimed at ensuring the safe use of any
new formulation meant for direct or indirect impact on food and feed safety, including
animal health and welfare, plant protection and plant health and nutrition. The
application process for EFSA approval are usually followed by aligning to pre-requisite
guidelines can be initiated by EFSA or national food safety authorities that are members
of the EFSA Focal Point network, For Ireland, The Food Safety Authority of Ireland (FSAI)
represents a statutory body responsible for ensuring food produced, distributed or
marketed in Ireland complies with food safety and hygiene standards, best practice
codes and legal requirements. Till date, EFSA has obtained 11 applications for claims on
health benefits of using fructo-oligosaccharides in nutraceutical/food formulations. The
commission authorises different health claims provided they are based on scientific
evidence and can be easily understood by consumers. The Committee concludes that
based on the data available at this time, fructans other than oligofructosyl-saccharose

10
should not be included in infant formulae and follow-on formulae10. From this projects
perspective, the validation of grass FOS for the development of nutraceutical
formulations for human consumption may require clinical trials and stringent
compliance measures which may not be in line with the scope of the farmer co-op’s or
SME’s venturing into commercialization of grass FOS products. However, the regulatory
measures to be followed for development of animal feeds are less stringent and
substantial research activities have established the benefits of FOS in animal diet (mainly
monogastrics)11. The regulatory measures to followed in development of animal feed
formulations are explained in the Subsections 2.2 and 2.3, respectively.

10
https://ec.europa.eu/food/sites/food/files/safety/docs/labelling_nutrition-special_groups_food-children-out199_en.pdf
11
Kozłowska, Izabela, Joanna Marć-Pieńkowska, and Marek Bednarczyk. "2. Beneficial Aspects of Inulin Supplementation as a
Fructooligosaccharide Prebiotic in Monogastric Animal Nutrition–A Review." Annals of animal science 16.2 (2016): 315-331.

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2.2. Protein feed for Monogastric animals

2.2.1. Introduction
The protein concentrates from grass are obtained by thermal hydrolysis of grass juice
after biorefining process. The protein concentrates are coagulated and dried for easy
transport and perspective use as feed in pig and poultry industries, respectively. Protein
concentrates thus obtained, replaces the need to extensively import soybean meal
(SBM) as a protein supplier and reduces the negative effects on the environment.
Getting feed from around the world causes transport emissions; soya is a particular
cause for concern Ireland, up to 90% of the soybean and maize products are imported
from Argentina, Brazil, and the USA. Almost 1.7mt of soya and maize genetically
modified (GM) products were imported into Ireland for animal feeds in 2017,
constituting approximately 50% of total feed imports12.

Several studies have substantiated the potential of grass proteins to be of the same
nutritional value from other sources. With import of genetically modified crops being
selectively scrutinized with stringent regulations by EU, use of grass protein feed
concentrates in pig feed which accounts to 8 % of Ireland’s agri-food output (2014) 13.
According to the FAO (Food and Agriculture Organization, 2018) 25% of the world's crop
harvests are contaminated with mycotoxins which can result in major economic losses.
There are certain feed ingredients that nutritionally are particularly good for pigs but at
certain times may have a higher risk of mycotoxins (e.g. soya hulls, maize, pollard etc.)
therefore it is important to understand the consequences of mycotoxins on pigs. In
comparison to ruminants, pigs are extremely sensitive to mycotoxicosis as mycotoxins
undergo a degree of degradation in the rumen which is not possible in pigs. Use of dried
protein concentrates from grass with negligible carbohydrate concentration can control
any fungal growth and curb any mycotoxin production in pig feeds.

2.2.2. Technical specification and product comparison with SBM

SBM is a high-quality protein feed (48% CP) with high energy content, high in by-pass
protein, good amino acid profile, being high in lysine but low in methionine, no limit on
inclusion, limited by protein requirement and price14. This section shall look in detail into
the performance of grass proteins against traditional soybean meal feed formulations
that have been used substantially in Ireland. Soybean meal is the by-product of the
extraction of soybean oil. Several processes exist, resulting in different products.
Soybean meal is usually classified for marketing by its crude protein content. High-

12
https://www.agriland.ie/farming-news/how-much-are-you-paying-for-your-concentrates/
13
https://www.teagasc.ie/media/website/animals/dairy/Whats_in_Grass_Todays_Farm_May2014.pdf
14
Bernard, John K. "Feed Ingredients| Feed Concentrates: Oilseed and Oilseed Meals." (2011): 349-355.

12
protein types are obtained from dehulled seeds and contain 47-49% protein and 3%
crude fibre (as fed basis). Other types of soybean meal include the hulls or part of the
hulls and contain less than 47% protein and more than 6% crude fibre. In solvent-
extracted soybean meals, the oil content is typically lower than 2% while it exceeds 3%
in mechanically extracted meals15. From Table 2, the crude protein contents (39.1 to
47.5%) of grass were noted to be within range of various soy bean meal processed via
various processing methods (37.56 to 55.62%).

Table 2: Product performance of Grass protein concentrates VS commercially available

soy-bean feed alternatives

Extruded Enzyme Soy

Full-fat Dehulled Nondehulled expelled treated Fermented Soy protein protein Grass protein
Product (%) soybeans SBM SBM SBM SBM SBM concentrate isolate concentrates

Dry matter 92.36 89.98 88.79 93.85 92.7 92.88 92.64 93.71 90

Crude
protein 37.56 47.73 43.9 44.56 55.62 54.07 65.2 84.78 39.1 to 47.5

Ether
extract 20.18 1.52 1.24 5.69 1.82 2.3 1.05 2.76 0.8 to 1.2

Carbohydrat
e and lignin 29.72 34.46 37.27 37.9 28.21 29.53 20.28 2 0.05 to 0.8

Ash 4.89 6.27 6.38 5.7 7.05 6.98 6.11 4.17 7.5 to 15

Table 2 lists out the nutritional values of various soy products used in pig feed which is
adopted from a report16. The grass proteins values were incorporated into it from
preliminary studies conducted at Grassa BV (project partner). These ingredients contain
little fat because they are co-products of oil production, and thus, the fat has been
removed. Dehulled soybean meal contains less than 2% fat on an as-fed basis, compared
with whole full-fat soybeans that contain approximately 20% fat. The comparison of
grass protein concentrate from this project should be made with soy meal because both
have the same properties along with matching amino acid profiles. it is quite evident
that the protein concentrations and more or less similar 47.73 % (for dehulled SBM) and
39 - 47.5 % (grass protein concentrates). The fat contents values are also similar in range,
whereas the fibre contents carbohydrate values are much lower. Several research works
have substantiated the benefits of various forage-based protein sources against SBM in

15
http://www.soymeal.org/ReviewPapers/SBMExceptionalProteinSource.pdf
16
Li, Zhongchao, et al. "Prediction of digestible and metabolisable energy in soybean meals produced from soybeans of different
origins fed to growing pigs." Archives of animal nutrition 69.6 (2015): 473-486.

13
animal feeds; the results from these studies majorly investigates the total body mass
changes which did not show any significant differences17.

2.2.3. Regulatory compliance measures

The European union ensures the verification of compliance with feed and food law,
animal health and animal welfare rules, outlines inspection and control procedures that
must be operated by Member States (Regulation (EC) No 882/2004)18. In order to
protect the EU from feeding formulations which pose a serious risk to animal or human
health, member states are required to set up a contingency plans/ and strictly adhere
to the regulations. Several regulations pertaining to hygienic safety requirements for
feed products are regulated by DAFM in Ireland. Within Europe, regulatory schemes
contain clear standards and requirements to guarantee safe feed and responsible work
methods throughout the chain which concerns production facilities, but also storage,
transport, personnel, and procedures. Integration of ISO standards, HACCP and other
parameters in production and use of feeds can deem the product to be fit for a GMP +
feed certification scheme. Several SME’s and in-farm feed producers have used the
GMP+ certification schemes for making the feed formulations which are guaranteed to
be safe and reliable.

In Ireland, the Health Products Regulatory Authority (HPRA), and the Department of
Agriculture, Food and the Marine (DAFM) is the Competent Authority responsible for
the transposition and implementation of European Union legislation on animal feeding
stuffs19. Within DAFM, the Feeding stuffs, Fertilizer, Grain & Poultry Division (FFGPD)
develop policy on animal feed, manage the negotiation, transposition and enforcement
of feed legislation and carry out official controls along the feed chain, to ensure that
these commodities do not present a risk to the food chain and that the required product
information (labelling) is given to customers. Qualified Presumption of Safety (QPS)
[EFSA], is a licensing protocol, similar to GRAS (Generally classified as safe) classification
used in USA (FDA) that has recently been introduced by the European commission. QPS

17
Kurrig, Magdalena, et al. "Replacing soybean meal by alternative protein sources in forage-based diets for growing bulls." EAAP
Scientific Series. Wageningen Academic Publishers, 2019. 1653-1662.
18
https://www.fsai.ie/uploadedFiles/Legislation/Food_Legisation_Links/Official_Control_Of_Foodstuffs/15Consol_Reg882_2004_01
Jan07.pdf
19
https://www.agriculture.gov.ie/media/migration/foodindustrydevelopmenttrademarkets/feedingstuffs/officialinspections/Animal
FeedControlsProgramme20122014.pdf

14
is a generic risk assessment approach applied to biological agents aiming at simplifying
risk assessments across different scientific panels and units 20.

20
Leuschner, Renata GK, et al. "Qualified presumption of safety (QPS): a generic risk assessment approach for biological agents
notified to the European Food Safety Authority (EFSA)." Trends in Food Science & Technology 21.9 (2010): 425-435.

15
2.3. Press cake as an alternative for grass silage

2.3.1. Introduction
Over 85% of farms in Ireland make silage each year. Grass silage accounts for 20-25% of
total annual feed per cow on well-run dairy farms, and up to 30% of total feed on beef
farms depending on the production systems in place21. Effects of feeding press cake
silage as replacement for high quality grass silage on milk production were analyzed at
Lyons farm, UCD. This would be essential in benchmarking press cake as an effective
alternative for grass silage in farm that undertakes grass biorefining process.

2.3.2. Technical specifications and performance of press cake against grass silage in
cattle milk production
The objectives of this experiment was to evaluate the effect of direct replacement of
traditional high quality grass silage (25% DM, 16% CP, 77% DMD) with press cake silage
(37% DM, 10% CP, 62% DMD) in the diet of dairy cows on milk yield and composition.
The results of these trials are elaborated in Deliverable reports 2.2 and 2.3, respectively.
In summary, there were no significant differences in milk yield and composition between
dietary treatments in this study. Results show that press cake silage can act as a partial
replacement for high quality grass silage in the diet of early lactation autumn calving
cows without having negative impact on milk yield and composition. The compositional
analysis of the feed trials and proximate analysis of the press cake against sillage samples
are shown in Tables 3 and 4, respectively. Although press cake cannot be a complete
alternative to silage- due to low protein, sugars, oils and VFA’s; partial replacement of
press cake (from biorefinery) could be viable solution in Ireland where there are ~50%
grassland are present and a cascade of various value added products could be generated
from it.

Table 3: Compositions of traditional silage and press cake feed mixes

Label Control Press cake

Grass silage 14 kg DM 5 kg DM
Press cake __ 9 kg DM
Concentrate 7.22 kg DM 7.22 kg DM
Soya bean meal 0.44 DM 0.44 DM
DMI (kg DM/day) 21.64 21.64
CP% 18.6 15.9

21 https://www.teagasc.ie/crops/grassland/quality-grass-silage/

16
 NDF% 38.3 45.6
UFL 18.94 17.33

Table 4: The average proximate analysis of silage vs press cake from 5 farms

Sillage Lyons farm (S) Press cake Avg 5 farms (PC)

DM 25.95 37.73
CP 16.5 10.11
DMD 77.2 62.6
ME MJ/kg 11.7 9.35
Sugars % 1.55 0.64
Ash 9.65 5.53
NDF 47.9 64.37
ADF 29.65 42.38
Lignin g/kg 30.6 61.87
OiL b 4.8 2.72
pH 3.75 4.19
Ammonia N 2.75 1.96
VFA's g/kg 26.15 10.38
Lactic Acid 122.3 31.74
PDIA 30 18.4
PDIN 97.55 59.77
PDIE 81.65 68.54
UFL 0.88 0.69

Rumen simulation technique (RUSITEC)- a well-established in vitro method to simulate

and to investigate rumen microbial processes and nutritional uptake within the cattle
rumen. This method was utilized by our partners at University College Dublin, for
determining performance of press cake constituted meals against traditional grass silage
meals. The feed efficiency values were quite comparable (no significant different at
p<0.05) to that of grass silage. The gas production values and methane production
values for press cake was lower on comparison with silage, however there were not any
significant differences (p<0.05).

2.3.3. Regulatory compliance measures

17
Roughly 5 million EU farmers raise animals for food production with a value of about
EUR 130 billion. Every year, they need approximately 450 million tons of feed for their
animals. Similar to Section 2.2.3, the guidelines for animal feed formulations are
directed by Food, Farming, Fisheries division of the European union, which ensures the
safety and quality of agricultural and food products, supports producers and
communities, and promotes sustainable practices. COMMISSION REGULATION (EU) No
691/2013 was the last updated document which summarizes the protocols and
directives set for sampling and analysis of performance of feed additions on farm
animals. In Ireland, DAFM-Department of Food Agriculture and Marine, oversees the
regulations and compliance measures to be followed within the country. DAFM
regulates the laws pertaining to the use of raw materials used in manufacture of feed
formulations including the additives used, the feed labelling protocols, pre-commercial
product validation including tests involved for undesirable products such as mycotoxins,
ochratoxins, and other pathogens. Recently, Regulation (EU)2020/354 has been
implemented in and should come into prevalence from December 2020, for curbing the
use of non-EU, GMO crops to be used in feed formulations. This would have a direct
impact in Ireland as the meat, poultry and dairy industry would be directly impacted by
this, as majority of the soy bean meals are presently being imported and would urgently
be required into looking at other alternatives for non-GMO based products.

Generally, prior to performing any sort of animal feed trials, the ‘Animal research and
ethics committee’ of an organization are usually examines the proposed feed
experiment and trails for scrutinizing the use of any substances which maybe deemed
as unfit or could have health implications of the animals health. The veterinary wing of
‘Health products and Regulatory Authority (HPRA), issues licenses for organizations and
farming Co-op’s for performing trials on animal feed and additives etc. The trails for pre-
commercial validation of press cake samples as fodder for cattle were performed
according to experimental license from the Health Products Regulatory Authority
(HPRA) under European directive 2010/63/EU and S.I. No. 543 of 2012.

18
2.4. Biofertilizer production & Digestates in Anerobic digestion

2.4.1. Introduction
Grass whey, a residue from the biorefining of grass has the potential to be marketed as
a replacement to synthetic fertilizer, thus making further inroads into Ireland’s
agricultural greenhouse gas emissions. In 2016, Ireland’s greenhouse gas emissions
were 61.5 million tonnes of carbon dioxide equivalent. This was a reduction compared
with the peak average annual emissions of 68.9 million tonnes in 2000-2004, but 3.6%
higher than in 2015. As farming has evolved in Ireland, more attention has been paid to
the nutrient content of fertilizers both organic and synthetic over the years in terms of
N, P and Potassium (K). The most natural and readily available fertilizer at farm level is
organic manure produced from animals. This slurry is generally spread back on the land
that was grazed in order to replenish N, P and K values. With the EU moving towards the
nitrates directive (91/676/EEC), it is imperative to look into wastes arising from grass
biorefinery- the whey after the value added products of proteins and FOS have been
extracted out, the slurry has the potential to be used as fertilizer to improve nitrogen
use efficiency on farm and ultimately reduce GHG’s and Ammonia emissions in Irish
agriculture.

2.4.2. Technical specifications of grass whey fertilizer.

Figure 4 summarizes a trial done by Carberry group on grass whey as a fertilizer
alternative on grassland, the trials monitored the growth of grass in comparison with
slurry which are generally used in general farming practices. The data indicates a
promising result; where whey-fertilized grass growth performance was shown to be
better than slurry. Detailed discussions on trials and results are elaborated in Deliverable
2.6, a few interesting excerpts are shown below.

The Nitrogen values of grass whey were noted to be higher that slurry. With high
concentrations of K and low concentrations of P, grass whey is suitable for using as a
fertilizer (similar to slurry or diluted slurry). The environmental hazards and foul odour
associated with slurry also makes the use of grass whey as an effective fertilizer
alternative. This approach enables a circular/ regenerative approach where residues
from grass biorefining process are recirculated back into farms for optimised growth.

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 Figure 4 :NPK values of slurry vs biofertilizer from grass

Biorefinery Glas also investigated an alternative approach where; grass whey residues
generated before and after extraction of FOS were tested for its potential as a feed for
anaerobic digestion processes. The two scenarios mentioned above gave perspectives
into the potential of grass whey to be used as an energy source via production of
biomethane from biorefining wastes. The results of these analyses have been
elaborated in D 2.6; results indicate that grass whey analyzed in both scenarios offer
good bio-gas potential and methane is a significant proportion of the overall gas volume.
This indicates the potential of whey to be used as a feed-stock in biomethane production
highly significant. It was also interesting to note that the residence time involved in
degradation of whey in both cases to biogas were very low ~5-6 days, this makes great
economic sense when compared with other commercial feed-stocks such slurry, which
takes a much longer time to digest. It was also recommended that the digestates
regenerated from these analyses had immense potential to be used as fertilizer as it was
shown to maintain optimal concentrations of Nitrogen/Potassium and Phosphorus, and
does not contribute towards acidification of soil with prolonged usage- due to the lack
of organic matter. Since both the streams of whey have had their protein constituents
removed, there are no possibilities of production of unwanted gases such ammonia or
hydrogen sulfide, which is a major advantage over using whey residues over slurry and
other proteinaceous side streams.

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2.4.3. Regulatory compliance measures
From June 2019, the EU Fertilizing Products Regulation (FPR) (EU) 2019/1009 has been
published in the Official Journal of the EU. FPR looks at an EU-wide end of waste criteria
for compost and digestate which can be used in organic fertilizers, soil improvers and
growing media. The new regulation states that; ‘There is also a need to make use of
recycled or organic materials for fertilizing purposes. Harmonised conditions for making
fertilizers made from such recycled or organic materials available on the entire internal
market should be established to provide an important incentive for their further use’22.
Promoting increased use of recycled nutrients would further aid the development of the
circular economy and allow a more resource-efficient general use of nutrients, while
EU’s dependency on nutrients from third countries. This is in line with the grass whey
residues obtained from this project and other second-generation biofuels derived from
lignocellulosic materials (including agricultural slurries) which complies with Ireland’s
Transition to a Low Carbon Energy Future, 2015-2030 (DCENR, 2015). Such stringent
regulations on the use of fertilizers for farmers has had its impact on curbing total use
within an Irish context. A study conducted by Teagasc between 2005 to 2015 -a period
when Ireland has been bound by the EU nitrates directive regulations governing fertilizer
use; nitrogen, phosphorus and potassium fertilizer application rates on grassland tended
to be between 11% and 16% higher when the survey began versus when it concluded23.
Furthermore, the study mentions that nitrogen, phosphorus and potassium application
rates for the main cereal crops for agri-environment scheme participants were – on
average – 15%, 10% and 3% lower, respectively over the study period. These norms have
helped hugely improve level of soil protection, reduce acidification of soil, and reduce
emissions of GHG’s and other toxic gases from soil.

With FPR in practice, it has opened the market for organic fertilizer producers to sell
their produce across the EU; there are more choices for farmers to choose from and the
environmental and health risks associated with the use of synthetic fertilizers are
mitigated.

22
https://www.agriland.ie/farming-news/new-fertilizer-regulations-adopted-for-eu-market/
23
https://www.agriland.ie/farming-news/survey-indicates-long-term-reduction-in-fertilizer-usage/

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 3. Conclusions
The Biorefinery Glas project has been able to demonstrate the potential of various
products which not only substantiates the potential of grass biorefineries in Ireland, but
gives farmers, SME’S and established agricultural conglomerates a perspective to
employ such systems within their businesses in future. The pilot/lab scale validation of
the performance of products mentioned in Section 2 not only gives us insight into how
it could be compared with commercial market standards, but the organic/non-GMO
nature of products originating from grass further justifies and falls within the various
environmental regulations associated with marketing it.

Looking at the regulatory framework required by farmers and SME’s to commercialize a

product into the market- the processes, costs and time involved are a major deterrent
for many small-scale manufacturers. The continuous changes in directives to a
regulation also pose a constant worry for SME’s to commercialize their product into
market. This being said, EU has streamlined several processes and have categorically
outlined the steps required for marketing a prototype/ food or feed formulation into the
market; for eg. Panel on Additives and Products or Substances used in Animal Feed
(FEEDAP) providing open consultation on a drafts & guidance on the
application/renewal of feed formulation licenses. In Ireland, Department of Agriculture
Food and The Marine over the years have also developed several schemes to help
manufacturers with technical assistance, providing information with various funding
calls aligning to the norms set by European Union. Overall, with majority of norms being
streamlined towards Zero GHG emissions targets; it is going to be a collective challenge
of effectively aligning all agricultural regulations in Europe. This is imperative especially
in Ireland, where the agro-food sector accounts 8% of the total gross value added and
provides almost 10% of the employment. More funding opportunities and campaigns to
provide information to stake-holders on regulatory compliance action plans are required
for this to be successful.

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