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Giuliano Regonesi -Microalgaex Innovation Center -Istanbul SPIRULINA IN

CATTLE FEED: ADVANTAGES

Preprint · May 2023

DOI: 10.13140/RG.2.2.10365.54246

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SPIRULINA IN CATTLE FEED: ADVANTAGES

BY Giuliano Regonesi – MICROALGAEX INNOVATION CENTER – ISTANBUL

May 2023

1 INTRODUCTION
In the coming years, the global demand for animal products is anticipated to surge as a
result of population expansion and shifting lifestyles in emerging nations. Consequently,
the sustainability of farms has become a pressing issue in light of evolving European
Union policies such as the Green Deal and Farm to Fork strategy. This report delves into
the utilization of spirulina algae as a substitute feed supplement for ruminants,
scrutinizing its impact on animal performance and environmental sustainability.
Spirulina, unicellular microalgae, is a protein-rich source that can utilize various raw
materials and wastes for growth and biomass production. As the basis of the food chain,
microalgae play a pivotal role in sustaining the Earth's oxygen levels and enhancing soil
fertility. Furthermore, our analysis encompasses an overview of current legislation
governing alternative feed production. With its eco-friendly and economical chemical
composition, spirulina presents a promising alternative for livestock feed. We discovered
that spirulina contains numerous biologically active substances that expedite the
digestive process and promote better nutrient supply, further solidifying its potential as
an alternative feed for farm animals. In conclusion, we advocate for additional research
into the use of spirulina algae as a supplement in ruminant diets to fortify sustainable
farming practices. The consumption of animal-derived products, including meat, fish,
eggs, milk, and their byproducts, endows humans with high-quality proteins that are rich
in vital essential amino acids crucial for our bodies. According to research conducted by
FAO, the global population is projected to reach 9.6 billion by 2050, with 70% residing
in cities and an average income that is double the current standards. This indicates a
sizeable increase in the demand for animal-derived products, underscoring their
indispensable contribution to global food security and nourishment.

Fig. 1: demand of animal-based products

The growth in global demand for animal protein will put our planet's precarious
resources in crisis, so the goal in recent years of looking towards the future is to produce
in a sustainable way and try to use alternative resources for animal feed. The livestock

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 1

sector is growing much faster than other rural sectors, especially regarding intensive
farming, and is responsible for emissions from land use and land use change, affecting
the increase in CO2 and producing a worrying percentage of greenhouse gases. Animal
husbandry is responsible for 37% of the methane produced (ruminant digestive system),
and 64% of ammonia which contributes to acid rain.

1.1 The pollution

Air
Air pollution is a grave concern, and methane emissions from the rumen of cattle only
add to the problem. Methane is classified as a powerful greenhouse gas with 23 times
the impact of carbon dioxide, making it a significant factor in climate change.
Interestingly, while 20-30% of the methane emitted comes from fossil sources, the
remaining 70-80% is produced through enteric fermentation, primarily from livestock.
In fact, cattle are responsible for a whopping 73% of all livestock methane emissions,
which contribute 15-20% of total anthropogenic emissions. To mitigate this issue, we
can focus on manipulating the rumen ecosystem by adjusting the diet, using alternative
food sources, and employing additives. With these viable solutions, we can hope to
reduce methane emissions and tackle air pollution head-on.

Fig. 2: GHG emission by sector (agriculture in evidence)

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 2

Fig. 3: Livestock and pollution

Soil
Regarding the environmental impact of zootechnical practices on soil, it is crucial to
understand that around 70% of the agricultural area and 30% of the non-glacial land
surface is being utilized for food production for this sector. This expansive use of land is
causing significant physical, chemical, and biological alterations, leading to soil
degradation. The most common degradation processes identified are desertification,
erosion, decreased organic matter, compaction, and salinization, which can diminish
the soil's primary functions. These changes are primarily caused by inaccurate
agricultural techniques, such as unbalanced fertilization, monoculture, grazing, and
deforestation. The adverse effects of soil erosion, mostly caused by trampling and
grazing, further exacerbate the situation by adversely impacting water levels and
depositing sediments on the surface. Thus, it is evident that soil pollution is a grave
concern and necessitates urgent attention.

Water
The interrelation between animal husbandry and water is a complex and nuanced issue.
The use of livestock manure as fertilizer and accompanying soil characteristics can result
in leaching and flow phenomena. Additionally, the nitrogen present in manure, sewage,
and agricultural fertilizers can lead to eutrophication. Nutrient loss is not solely

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 3

determined by quantity but is also influenced by various factors including soil and
climatic conditions, methods and timing of distribution, and soil characteristics. Nitrogen
in animal manure exists in both organic and inorganic forms, with the former exhibiting
limited movement in the soil, while the latter - particularly ammonium ions and nitrates
- are susceptible to volatilization and leaching. This highlights the importance of
carefully considering the interplay of agricultural practices and environmental factors to
address water pollution."

Biodiversity
In recent years there has been a decrease and even the disappearance of some species
and related habitats, ecosystems and genetic heritage. The factors that negatively
influence biodiversity are mainly land use change, climate change, the introduction of
invasive species and pollution.
Maintaining biodiversity is essential for human life, well-being, and the long-term
sustainability of agriculture and animal husbandry.
Biodiversity concerns both wild and domestic species but it can also concern plant
species. Livestock farming contrasts with biodiversity because, first of all, there is
competition for surfaces between domestic animals and wildlife and secondly we find
land use change. Furthermore, there have been losses of genetic resources due to the
use of cosmopolitan breeds at the expense of autochthonous ones.

Feeding
The notion of diet has gained widespread attention in recent years, particularly with
regard to adherence to the four principles of the 4Rs: reduction, reuse, revaluation, and
research. Reduction entails limiting the competition between human and animal food
sources by exploring alternative energy and protein sources. Reuse involves repurposing
food waste, while revaluation entails transforming agri-food and industrial by-products
into functional foods. Finally, research is key to understanding the characteristics of
foods to optimize their usage.

To ensure high-quality animal products, it is essential to source new, nutrient-rich food

and optimize land and water use. Currently, the feed industry in Italy largely relies on
soy flour for its protein sources. The EU's "European Protein Plan" aims to increase
vegetable protein properties and explore alternative sources like insect-derived flours.
Industrial by-products and biofuels, including distillates, panels, and extraction flours,
are also viable options.

Additionally, spirulina algae have gained credibility as a novel food. Experimental trials
have proven its effectiveness as a protein source for chickens, pigs, rabbits, and
ruminants. It is important to note that spirulina algae fall under the novel food legislation
for the European Union, which regulates all non-traditional food sources.

As such, continued research and exploration of alternative food sources are necessary to
ensure a sustainable and nutritious food supply.

2 TARGETS
The livestock supply chain is affected by global changes that bring both short and long-
term effects. This paper aims to analyze alternative feeds, specifically spirulina algae, to

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 4

determine the best method of use. The sustainability of zootechnical farms holds
significant importance economically, socially, and environmentally. Shockingly, 84% of
feed products are not edible for humans. With an estimated 9.6 billion inhabitants
predicted by FAO by 2050, it is crucial to optimize resources effectively. Although
spirulina algae are not yet fully understood, it is rich in proteins and a potential alternative
food source. Through an in-depth analysis of various studies concerning its use, this
paper examines the addition of spirulina algae to cattle rations. Despite having a high
production cost, spirulina algae is an excellent solution for land use as it grows and
develops in all aquatic environments, leaving croplands for human food untouched.
Nonetheless, there are limited articles available on this subject. Our findings support the
need for further research on the feasibility and cost-effectiveness of spirulina algae as a
sustainable alternative feed.

3 ALTERNATIVE FEED FOR RUMINANTS: NUTRITIONAL VALUE, PRODUCT

QUALITY AND ENVIRONMENTAL ASPECTS
In the quest for sustainable livestock systems, there is a growing interest in alternative
feeds for ruminants and their potential impact on nutritional value, product quality, and
environmental aspects. Researchers are exploring ways in which fruit and vegetable
residues can be incorporated into the animals' diet, even though their preservation is
more challenging compared to traditional byproducts from sugar, starch, alcohol, and
oil production. According to Shader et al. (2015), a shift towards an animal diet that
comprises solely food industry byproducts coupled with a reduction in human
consumption of animal-origin products and grazing could lead to a marked reduction in
pollution. Specifically, greenhouse gas emissions, nitrogen and phosphorus loading, and
land and water use could decrease up to 46%.

Fig. 4 Bilancio proteico Globale

An adequate study of the diet with supplements and food choices to modify the ratio of
ruminants has good potential both to improve total digestibility and to increase
performance and at the same time decrease methane emissions. Intensive agriculture is
a significant source of nitrogen, in fact, about 50% of that is brought to crops as fertilizer
is not used by plants, but is lost in the environment as ammonia, nitrate and nitrous

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 5

oxide. Nitrogen-fixing legumes can be a valid ecological and sustainable alternative for
the diet of ruminants, thus reducing nitrogen losses and optimizing dietary nitrogen.
For the use of alternative feeds, it is important to know the value of new feeds, but also
the responses of animal production and the costs of new feeds compared to conventional
ones. The environmental footprint also plays a fundamental role in choosing the use of
new products.
Wood is the most abundant carbohydrate in the world, but untreated it is difficult to use
by ruminants due to the presence of bonds between cellulose and lignin, even if its
usability can be improved with various processing methods. Globally, the advantages of
unicellular proteins (bacteria, yeasts, fungi, microalgae) and aquatic biomass (algae,
duckweed) compared to terrestrial crops are the independence of arable production and
weather conditions. The chemical composition of these new feeds varies depending on
the species and growing conditions. For example, microalgae have good potential both
as lipids and as protein supplements in ruminants. Algae produce bioactive substances
that have the potential to reduce methane emissions.
The advantages of seaweed production are the independence of seed production, non-
dependence on climatic conditions, high production and continuous harvesting.
However, the cultivation, harvesting, storage (especially drying) and application of these
new types of feed on a large scale require further studies to lower production costs and
make them competitive with existing products in the market. In the long term, microalgae
have the greatest potential to become the leading sources of vitamins and proteins
worldwide.

FOOD NEED FOR WATER ENVIRONMENTAL ASPECTS

Rapeseed High need nitrogen to get high yields
Soy High The day length and low temperatures limit
the potential for yield and expansion
Grain High need nitrogen to get high yields
Lupins, peas, beans High The high rumen degradability of proteins
and their unbalanced amino acid profile can
increase nitrogen production
Legume forage High Due to the low content of long fibres, they
can limit methane emissions
High May improve N and P use efficiency if diet
Hemicellulose exceeds in these nutrients
Leaves of tropical trees High In certain species they mitigate the
production of methane
Microalgae Low Based on the components they possess
there is a decrease in production of
methane.
Algae None Nutrients are absorbed from the marine
environment, early harvesting can
compromise the balance Coastal
Duckweed Low They recover nutrients from wastewater

Table 1. Potential environmental effects of different types of foods

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 6

4 PHYTOPLANKTON
Derived from the Greek words phyto (plant) and plankton (made to wander or drift),
phytoplankton are microscopic organisms that live in watery environments, both salty
and fresh. Some phytoplankton are bacteria, some are protists, and most are single-celled
plants. Among the common kinds are cyanobacteria, silica-encased diatoms,
dinoflagellates, green algae, and chalk-coated coccolithophores.

They inhabit the watery depths and play a vital role in our planet's functioning. These
single-celled photosynthetic powerhouses dwell near the water's surface, basking in the
sun's rays and converting carbon dioxide into oxygen-rich carbohydrates. As nature's
primary producers, they form the foundation of the oceanic food chain.
The euphotic zone, the illuminated realm where sunlight penetrates enough to fuel
photosynthesis, determines the reach of these remarkable beings. Though the clarity of
the water affects its thickness, this zone typically stretches up to a mere 200 to 300 meters
(or 600 to 900 feet) in a vast ocean that plunges 4,000 meters (13,000 feet) deep.

Two distinct camps of phytoplankton emerge. The first group boasts grandeur in the form
of single-celled algae, protists that possess sophisticated eukaryotic cells akin to
protozoans. Most frequently found near coastlines, these algae, including the esteemed
diatoms, occasionally experience explosive population growth known as blooms,
responding to fluctuating seasons and the availability of nutrients like nitrogen, iron, and
phosphorus.

In the other corner exist the humble giants—the photosynthetic bacteria. Despite their
minute size, some measuring only a micron across and numbering in the hundreds of
thousands per tablespoon of seawater, these invisible powerhouses dominate the sea.
Unfathomable by nets, these organisms languished in obscurity until the 1970s, when
scientific advancements finally laid their world bare. Today, we understand their central
role, as they account for half of the ocean's primary productivity and stand as the most
abundant life forms within the watery abyss. This group also includes cyanobacteria,
believed to be some of Earth's oldest organisms and the origin of plant cells' chloroplasts,
where photosynthesis takes place.

What makes these minuscule organisms so significant? Phytoplankton occupy an

essential niche in our world, providing us with a vital source of oxygen. In fact, they
generate approximately half of the atmosphere's oxygen, holding equal standing with
land plants. Furthermore, they form the bedrock of virtually every oceanic food web,
enabling the thriving existence of countless organisms that roam the sea.

Delving deeper into the significance of these remarkable beings, phytoplankton partakes
in the intricate dance of Earth's carbon cycle. Through photosynthesis, they transform
inorganic carbon present in the atmosphere and seawater into organic compounds,
playing an indispensable role in keeping our planet's carbon equilibrium in check. When
they meet their ultimate fate and descend to the depths of the ocean, they transport the
atmospheric carbon with them, effectively influencing the climate system.

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 7

Iron, or rather the scarcity of it, often serves as a barrier to phytoplankton growth.
Consequently, scientists engage in ongoing discussions around the prospect of fertilizing
large swaths of the ocean with iron to stimulate phytoplankton blooms, in turn facilitating
the greater transfer of carbon from the atmosphere to the ocean depths.

Phytoplankton also proves to be a key player in the intricate biogeochemical cycles that
take place beneath the waves. They diligently absorb, transform and recycle various
elements required by other organisms, ensuring the continuous flow of necessary
nutrients. In particular, photosynthetic bacteria excel in the nutrient-free expanses of the
open ocean, finding and releasing valuable vitamins and micronutrients that support
other marine life.

Yet, the impact of certain phytoplankton extends beyond ecological marvels. Dense
blooms of specific species can wreak havoc on coastal waters, depleting oxygen levels
and suffocating fish and shellfish. Additionally, particular phytoplankton unleashes
toxins that pose risks to humans and animals alike. Exposure to these toxins or
consumption of contaminated shellfish can lead to illness or even tragedy for
unsuspecting individuals and even our magnificent oceanic leviathans, the whales.

In light of these profound insights, it becomes abundantly clear that the study and
understanding of phytoplankton lie at the crux of our collective responsibility. These
humble yet powerful organisms fuel our planet, support thriving ecosystems, and actively
shape our climate system. Grasping their intricate mechanisms and delicate balance
ensures our ability to preserve and protect the wonders that reside beneath the waves.

Fig. 5: Phytoplankton are extremely diverse, varying from photosynthesizing bacteria (cyanobacteria), to
plant-like diatoms, to armor-plated coccolithophores (drawings not to scale). (Collage adapted from
drawings and micrographs by Sally Bensusen, NASA EOS Project Science Office.)

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 8

5 ALGA SPIRULINA

Fig. 6 Spirulina algae

Spirulina algae, also known as Spirulina Platensis, is a remarkable source of protein, rich
in essential nutrients, and a well-studied unicellular microorganism. Capable of utilizing
an array of raw materials and waste, Spirulina exhibits a versatile growth pattern to
produce protein-rich biomass. Though protein contents differ according to strain and
species, allergens, heavy metals, pesticides, and toxins are potential hazards, particularly
when grown on contaminated substrates. However, even in the absence of readily-
available nitrogen, the degradation of nucleic acids and their derivatives in the rumen
provides a renewable source of carbon and nitrogen for bacterial growth. The crucial
stages of Spirulina protein production include preparation of the culture medium,
photosynthesis, harvesting, as well as washing, cell destruction, extraction, and protein
purification. While production costs remain a challenge, the ever-growing need to
safeguard the environment, and intensify nutrient use and resource management,
coupled with the possibility of protein scarcity, suggest a promising future for Spirulina
as an economically-feasible protein source. Microalgae, like Spirulina, are small aquatic
organisms that solely utilize light energy to synthesize a host of vital substances,
including antioxidants, proteins, vitamins, and omega-3s, which can greatly benefit both
humans and animals. Spirulina, with its helix-like, unbranched filamentous structure,
dates back 3.6 billion years and represents the oldest known form of photosynthetic life.
Found mainly in subtropical freshwater environments, the alkaline pH of 9 to 11 is
optimal for its growth. Spirulina's balanced biochemical composition lends it an array of
beneficial properties, making it a highly-regarded "green food" and the subject of recent
nutritional studies.

Proteins: contains a high percentage of proteins between 55 and 70% of dry weight;

• Essential fatty acids: it has a high concentration of PUFA and 2% of total lipids, it is
rich also in linoleic acid.
• Vitamins: contains vitamins B1, B2, B3, B6, B9, B12, C, D and E;
• Minerals: it is a rich source of potassium, calcium, copper, iron, magnesium,
manganese, phosphorus, selenium and sodium;
• Photosynthetic pigments: contains chlorophyll-a, xanthophylls, β-carotene,
zeaxanthin, canthaxanthin and phycobiliproteins.

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 9

 Nutrients % composition Details
Proteins 65 8 essential amino acids
Carbohydrates 15 10 non-essential amino
acids
Lipids 6 7 types of acids (alpha
linoleic..)
Vitamins 0.75 B1, B2, B3, B6, B9, B12, B7,
D, B5
Minerals 8 Micro and macro elements
Carotenoids 346 mg/100g Alpha-Carotene, Beta-
carotene

Table 2. Spirulina Platensis nutrients

Spirulina stands out as a rich source of highly bioavailable proteins, particularly

biliproteins, comprising 65% of its nutrient composition. The high degree of utilization
of these proteins, around 85%, makes it a valuable alternative to conventional proteins.
As an animal feed, microalgae exhibit a nutrient profile that is highly variable, making it
flexible in multiple applications. Compared to soybean and rapeseed meal, microalgae's
amino acid composition is similar, except for a relatively lower amount of histidine. It's
worth noting that histidine is the first limiting amino acid that affects milk production on
silage, grass, and grain-based diets. The protein degradability of numerous microalgae
species is thought to be higher than that of oilseed rape.

Compared to other microorganisms, spirulina's murein shell, instead of a cellulose wall,

makes it easier to digest without any cooking requirement. Animal protein digestion
leads to the formation of metabolic waste, which is not the case with spirulina protein
digestion. Blue-green microalgae are vital components of the food chain that generates
over half of the world's oxygen. Unfortunately, pollution destroyed many natural sources
of spirulina, which is now cultivated in pristine areas. The quality of the seaweed
depends on its origin and processing method, significantly impacting its nutritional
quality.

Spirulina supplementation has promising properties such as antioxidant,

immunostimulant, anticancer, anti-inflammatory, cardioprotective, antidiabetic,
hepatoprotective, antimicrobial, neuroprotective, and energizing. The Food and
Agriculture Organization has classified spirulina as an ideal dietary supplement that
provides advantages such as antioxidant and energizing properties and supplement
mineral and vitamin deficits.

The effects of spirulina algae are given by:

- Phycocyanin: it is a pigment present in high concentration in blue algae which has the
effect to eliminate hydroxyl radicals.
- Polyphenols: these are useful molecules, especially in the prevention of chronic
diseases digestive.
- Betacarotene: it has anti-radical action.

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 10

In order to enhance the taste and appeal of microalgae for animal consumption, various
methods of processing, such as pelletizing, can be applied. Additionally, the on-site
production of microalgae via closed ponds or photobioreactors integrated with the
watering system of livestock can lower the energy demands associated with the drying,
storage, and transport of feed. This approach has the potential to facilitate the cultivation
of microalgae as a viable and sustainable option for extensive agriculture in the
foreseeable future.

5.1 Production Phase

The production phase of spirulina cultivation boasts numerous advantages when
compared to traditional crops. With a whopping 60% protein on a dry basis and rapid
growth, this alga delivers protein yields that are 20 times higher than soybeans and 40
times higher than corn. Furthermore, it can be grown on saline lands, in small quantities,
and with minimal water loss due to evaporation. The use of only 2100 l/kg of water for
every kg of protein produced is remarkable, and the culture medium can be recycled.
Spirulina production currently thrives in 22 nations with an annual world output of
around 3000 tons. Typically, high yields are grown in deep water basins, which span
from 1000m2 to 5000m2 and have a depth of 15 to 18 cm. As for the growth stages,
microalgae has mainly four.

1. Adaptation phase (lag phase);

2. Logarithmic growth phase (log phase);
3. Stationary phase;
4. Declining phase.

During the initial phase, microalgae exhibit metabolic activity that aids in cell
enlargement and synthesis of crucial enzymes and compounds. This stage is critical in
preparing for subsequent phases wherein cell division and reproduction take place. The
log phase follows, characterized by an exponential increase in cell count. The stationary
phase then follows suit, with balanced rates of cell division and death leading to stable
population size. Ultimately, the declining phase sets in as the number of cells that
disintegrate exceeds that of reproducing cells, thereby halting growth. This could be
attributed to high algal concentration, nutrient depletion, or the buildup of toxic
metabolites.

Fig. 7 Spirulina algae growth

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 11

5.2 Production method

For the production of microalgae there are essentially two types of method:

• In open pond;
• In photobioreactors.

The first method, extensively studied,

encompasses a variety of open basins,
comprising both natural and artificial water
sources. The natural water options include
lakes, lagoons, and ponds, while the artificial
water sources comprise basins and
containers. The most commonly used systems
encompass broad but sufficiently shallow
basins, tanks, circular tanks, and raceway
ponds. Raceway ponds, in particular, exploit innovative technologies and knowledge
concerning the treatment of wastewater. These ponds are basins that are excavated into
the ground and waterproofed with plastic membranes. They boast a depth that can
extend up to 20-30 cm and a section reserved for mechanical blades that enable
exchange with the atmosphere. Given that the oxygen that spirulina algae produce
during photosynthesis must be removed to enable optimal growth, this type of pond
proves immensely beneficial. The sole drawback of this methodology is its need for
extensive areas that could alternatively be utilized for agricultural purposes. In contrast,
closed basins offer better environmental control albeit at high management costs. These
basins remain covered by greenhouses that facilitate cultivation through different seasons
and extend growing times. In the case of heated greenhouses, producers can
contemplate year-round production. Additionally, this system enables increasing the
quantity of carbon monoxide and consequently, accelerating the growth rate of the
algae.

Fig 8: open raceways

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 12

Fig. 9: closed pond

One effective methodology employed in algae growth is the utilization of

photobioreactors - advanced cropping systems designed to maximize microalgae
growth, as well as that of cyanobacteria and photosynthetic bacteria. Photobioreactors
can be either closed or open, with the former requiring state-of-the-art technology to
regulate parameters for optimal output. Closed photobioreactors function as enclosed
structures, shielding the algae from atmospheric pollutants and ensuring a controlled end
product. These structures come in various types including, but not limited to, tubular,
flat panels, and vertical columns, each with unique functionalities.

TUBULAR It is one of the most suitable for large

masses, they consist of a glass or plastic
tube and the recirculation of the algae takes
place both with the pump and with air
transport, they perform well if used
outdoors.
Flat panels They have a large lighting surface, consist
of a flat plate and transparent material for
maximum use of solar energy, there is a
good accumulation of oxygen even if less
than the tubular ones.
Vertical column They are compact, low cost and easy to use.
From these we obtain a good production of
algal mass.

Table 3. Different types of photobioreactors

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 13

 - Large lighting surface;
- Suitable for outdoor crops;
- Good biomass production;
- encrustations;
- Abundant wall development;
- Relatively cheap

Fig. 10: Tubular photobioreactor

- Wide illuminated surface;

- Suitable for outdoor crops;
- Good algal productivity;
- Easy to clean;
- Low accumulation of oxygen;
- High number of compartments;
- Difficulty in controlling the
temperature.

Fig. 11: flat photobioreacto

- High mass transfer;

- Good mixing with minimal effort;
- Easy to sterilize;
- Easily tempered;
- Reduced photo inhibition and
photos
oxidation;
- Small surface of
lighting;
- Use of sophisticated material.

Fig. 12: Vertical column photobioreactor

The successful cultivation of microalgae requires careful consideration of several key

factors. These include limiting the depth of the water to allow for optimal solar radiation

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 14

penetration and the removal of excess oxygen to inhibit the biological processes of
spirulina. Cooling of the culture liquid can be achieved through spontaneous
evaporation, although continuous monitoring of the process is necessary to ensure
proper thermal lowering. Closed basins are a superior choice, offering energy efficiency
and a favourable cost-benefit ratio compared to photobioreactors which require more
advanced technologies. Mixing the crop can be achieved through manual or wind-
powered systems while gauging maturity through the observation of its dark green hue.
Harvesting involves filtration on cloth and sun drying. While the cost of production lies
largely in the growing period, this can be offset by the use of available waste effluents.
The use of plastic lining materials such as PVC reduces the manufacturing cost in large
production plants. Constant monitoring of vital parameters is critical in maximizing
production.

5.3 Harvesting

The liquid contained in the tanks is conveyed to a filtering unit which allows separating
it into two distinct streams:

- The wet algal mass, where there is the greatest concentration of the culture;
- The filtered liquid is put back into the tanks as it is still rich in salts and nutrients useful
for the growth of the microalgae.

After harvesting the algal mass, it can be processed in two different ways. Either it is
collected in food cloth bags and subjected to a pressing to reduce its humidity to a
minimum, then the seaweed paste is inserted into a drawing machine which allows you
to obtain spaghetti with a small diameter, about 1.55 mm, this allows dehydration at low
temperatures which will further reduce the humidity of the product without deteriorating
its nutritional properties. The seaweed noodles are placed in food looms and sent to a
drying cell, which is fed with heat from the biogas plant.
Or the algal mass is sent directly to drying, carried out at 50° for a few hours, once it is
dried it is pulverized and bagged.

6 ACCUMULATIONS OF HEAVY METALS IN SPIRULINA

The accumulation of heavy metals in Spirulina poses a significant threat to both the
ecological and nutritional spheres. While some heavy metals like Copper, Iron,
Manganese, and Selenium do play important physiological functions in living organisms,
a majority of them are biological toxins even at low concentrations. Heavy metals have
found extensive applications in various working fields, leading to their release into both
surface and underground water resources, which pose a considerable ecological threat.

Within biological systems, heavy metals can disrupt several cell organelles and
membrane components, including mitochondria, lysosomes, endoplasmic reticulum,
and nuclei, ultimately leading to enzyme dysfunction by binding with thiol groups
(Wang e Shi,2001; Ayangbenro e Babalola,2017).

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 15

One effective method to remove and recover metals is microbial remediation, where
microorganisms use bioabsorption, bioaccumulation, biomineralization, and
biotransformation to restore environmental balance.

Bioaccumulation, a crucial biological phenomenon where metals are taken up through

several steps, including adsorption, precipitation, complexation, and active transport,
presents a promising approach for metal accumulation. Spirulina platensis cyanobacteria
have demonstrated their capability to accumulate metals from the atmosphere, making
them a suitable candidate for metal accumulation from single and multi-component
systems. (Zinikovscaia et al., 2015, al.2016).

In conclusion, the increasing accumulation of heavy metals in Spirulina demands our

attention, and environmentalists and scientists must come together to address this
ecological threat professionally.

Fig. 13: Dried spirulina and tablets

The accumulation of toxic substances in the environment is a major concern faced by

countries worldwide. Among these substances is cadmium, a malleable silvery-white
metal that can be found in trace amounts in various elements such as the earth's crust,
air, and water. It is named after the city of Cadmos where it was first extracted in ancient
times. Cadmium easily reacts with other elements, forming toxic compounds and is
rarely found in its pure form. Cadmium sulfate and chloride are two of the compounds
that dissolve most easily in water while cadmium sulfide is often the primary mineral
form. Cadmium is released into the environment from both natural and industrial
sources, including non-ferrous metal extraction, refining, processing, battery and paint
production, the use of phosphate-based fertilizers, and the incineration and disposal of
waste.
When present in the air, cadmium can travel long distances and settle on the ground or
surface of the water, typically accumulating in high levels in industrial areas, hazardous
waste sites, busy roads, and agricultural fields. In agricultural fields, cadmium binds to
the organic matter in the soil and is subsequently absorbed by crops, eventually entering
the food chain. In surface waters, cadmium can contribute to bioaccumulation.

While cadmium was once used only for tanning leather and as a pigment in limited
quantities, it has now found use in various fields. The significant impact of cadmium on

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 16

the environment reinforces the importance of proper waste management and safety
practices in its handling and disposal.

1. Rechargeable batteries; (being a disposable product, it constitutes the majority of

pollution)
2. Dyes;
3. Coatings and Platings;
4. Stabilizer in plastic materials;
5. Alloys with other metals for solders.

Due to its low acceptable exposure limit, overexposures can occur even in situations
where no traces of cadmium are found.
The binding mechanism between heavy metal ions and the toxic effect caused by them
differ in different microbial species. Usually, these ions are transported inside the cell
through protein transporters, the concentration of metal ions transferred in it also
depends on the amount of ions that are located near the transport channel.
The mechanism of bioaccumulation of heavy metal ions by microorganisms is not yet
fully known, Chojnacka and Wojciechowski described this mechanism in two phases:
an active phase and a passive phase. In the first phase, there is a preliminary and rapid
binding of the metal ions to the wall of the spirulina algae, in the second phase instead
we have a slower transport through the cell membrane.
It is very important to know that the growth of algae cultivation is related to the
dependence on light and many models derive from a combination of light attenuation
phenomena and a light response curve based on the Beer-Lambert law and an equation
Monod type. Kim et al. and Bagus applied the light-dependent Monod equation for
cyanobacteria in the photobioreactor. The bioaccumulation experiments are conducted
at optimal light intensity and therefore it is not possible to apply the models based on
the light intensity deviation.

growth yield

if we indicate with ∆x the increase in biomass resulting from the use of the quantity ∆s
of the substrate, it is defined as growth yield or biomass yield and is indicated with Y x/s
or Y x,s
Δ𝑥 𝑑𝑥
lim = = = 𝑌 = 𝑌𝑥/𝑠
∆"#$ Δ𝑠 𝑑𝑠
considering that the biomass produced and the substrate consumed vary in the opposite
direction, we have

𝑑𝑥
𝑌 =−
𝑑s
It has been widely demonstrated that biological materials are able to adsorb heavy metals
from aqueous solutions, especially if the metals are present in concentrations lower than
1 mg/L. According to the World Health Organization, the cadmium concentration in
unpolluted natural waters should be less than 0.001 mg/l. Many researchers have

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 17

reported the bioabsorption of cadmium by live and immobilized cells of Spirulina
Platensis, the bioremediation potential of Spirulina has also been investigated with almost
all heavy metal ions and other toxic compounds.
The studies are carried out in an agitated flask under restricted conditions, while the
commercial production of Spirulina biomass is carried out using large ponds.

7- STANDARDS AND REGULATIONS

As regards the European legislation on the use of algae in zootechnical feed, there is no
real Community regulation. By law, the term marine algae refers to multicellular marine
algae, phytoplankton and microalgae. With regulation (EU) 2015/2283, it is defined that
the seaweed product and its derivatives are new products for which it will take time to
leave room for experimentation to determine the doses that are possible when used as
food additives for production purposes. Important, however, is the recommendation (EU)
2018/464 which regulates the monitoring of metals and iodine in seaweed, halophytes
and seaweed-based products. This COMMISSION Recommendation (EU) 2018/464 of
19 March 2018 – concerning the monitoring of metals and iodine in seaweed,
halophytes and seaweed-based products states that: “Member States, in collaboration
with operators of the food and feed sector, monitor during the years 2018, 2019 and
2020 the presence of arsenic, cadmium, iodine, lead and mercury in seaweed,
halophytes and seaweed-based products. Monitoring should include edible halophytes,
including Salicorna europaea and Tetragonia tetragonoides, as well as a wide variety of
marine algae species, reflecting consumption habits and feed uses [...], to allow for
accurate estimation of the exposure. Occurrence data should also be collected for
seaweed-based food additives [...].” “The species or numbers of additives should be
indicated and whether fresh, dried or processed products were analyzed. If possible, the
origin of the products (wild or cultivated), the date and place of collection and the
proportion of seaweed that has been analyzed should also be specified, providing any
information on the label of the final products. “Monitoring data should be reported to
EFSA on a regular basis [...].” “In 2006, the Scientific Committee for Food established an
upper iodine intake limit of 600 μg/day for adults and 200 μg/day for children aged
between 1 and 3 years. It clarified that the ingestion of seaweed products with a high
iodine content, especially dried products, can lead to a dangerously excessive iodine
intake if such products contain more than 20 mg of iodine per kg of dry matter.

8 INFLUENCE OF SPIRULINA ALGAE IN THE PRODUCTION OF METHANE

The rumen is the first stomach of ruminants and provides microorganisms with a suitable
habitat for growth. Inside this first stomach, we find microbes of various kinds: protozoa,
anaerobic bacteria, and anaerobic fungi, derived from the degradation of the feed.
Microorganisms help degrade lignin and cellulose materials, generating proteins,
vitamins, and short-chain organic acids. The acids produced in the rumen are acetic,
propionic, and butyric and contribute to animal nutrition.
Rumen microorganisms have evolved over time, thanks to the use of complex
biopolymers of plant cell walls such as cellulose, hemicellulose and pectin. This
degradation involves 21 different enzymatic activities and a multi-enzymatic
extracellular complex called cellulosome has been isolated, this has been identified from
anaerobic bacteria as a factor that binds bacterial cells to cellulose particles minimizing
losses. Another enzymatic activity of great interest in the rumen is the fermentation of

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 18

carbohydrates by anaerobic fungi, which digest almost all the structural carbohydrates
in plant cells, this activity is produced by the vegetative phase of the zoospores, the
products obtained are lactate, acetate, hydrogen and carbon dioxide. The rumen fluid
was tested for the degradation of cellulose and lignin residues.
Studies have been performed that evaluated the hydrolysis of residual microalgal
biomass as feed for ruminants. These studies suggest that lipid-extracted microalgae are
highly digestible by rumen microorganisms, indicating the potential of these
microorganisms to degrade microalgae cells as a pretreatment for biofuel production. In
another study, Spirulina platensis was included in the drinking water intake of cattle,
showing an increase in blood serum urea concentration, suggesting the prompt
degradation and release of microalgae proteins bypassing the rumen. In the following
table, we have the main enzymatic activities necessary for the hydrolysis of microalgae
and the microorganisms that carry out these activities in the rumen.

different enzyme activity Bacteria Fungi Protozoa

substrate
polymers present
in the microalga
cellulose, Cellulase, Butyrivibrio Neocallimastix
hemicellulose, Hemicellulase, Fibrisolvens Frontalis
pectin Pectinase Ruminococcus Neocallimastix
albus patriciarum
Ruminococcus Caecomyces
flavefaciens communis
Fibrobacter Piromyces
Succinogenes communis
Starch Amylase Ruminobacter
amylophilus
Selenomonas
lactilytica
Lachnospira
multiparus
Pectin Pectinase Lachnospira
multiparus
Protein Protease Butyrivibrio Entodinium
fibrosolvens caudatum
Entodinium
simplex
Chitinase Clostridium
tertium
Lipids Lipase Anaerovibrio
lipolytic
Table 4. Algal substrate polymers

9 THE USE OF SPIRULINA IN THE DIET OF CATTLE

The analysis of the bibliography available on the subject on this document has
highlighted few contributions referring to livestock systems of different types.

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 19

The results of the scientific articles are highlighted in the table below.

Reference Farming Metodhs main results

system
▪Holman, A. E. O. Malu- Aduli Dairy cows 200 g of spirulina Increased milk
(2012). Spirulina as a livestock grazing in per day production, but
supplement and animal feed. Australia few studies to
Jurnal of Animal physiology and compare
animal nutrition, 615-623.
▪ E. Manzocchi- B. Guggenbhul- Grazing 15-30 g/day of Reduction of
M. Kreuzer- K. Giller (2020). dairy cows spirulina rumen
Effects of the substitution of fermentation,
soybean meal by spirulina in a modification of
hay-based diet for dairy cows on milk sensory
milk composition and sensory characteristics
perception.
Journal of Dairy science, 11349-
11362.

▪ J. Kulpys- E. Paulauskas- V. Dairy cows Integration with Improved

Pilipaviþius- R. Stankeviþius in 200 g of spirulina production, fat
(2009). Influence of Lithuania, content, protein
cyanobacteria Arthrospira grazing and even lactose
(Spirulina) platensis biomass
Bovine da latte in Lituania, al
pascolo
Integrazione con 200 gr di
spirulina
Miglioramento di produzione,
contenuto di grasso, proteine e
anche lattosio
additives
body
lactation
biochemical milk indexes.
Agronomy Research 7, 823-835

Table 5. Articles analyzed

LITTERATURE - EXPERIMENTS IN AUSTRALIA (Holman et al. 2012)

In Australia, the demand for dairy products is increasing due to the search for protein
products and consequently, there is also an expansion of the dairy products market. As
per tradition, Australian farmers prefer pasture as a source for feeding cattle, the big
disadvantage in using this technique is that the energy value of the grass in the pastures
is not sufficient to cover the needs of the dairy for the production of some milk.
Furthermore, the fat content in the milk of cows kept exclusively on pasture is of very
little value. Therefore, the alternative is supplementation with nutrient-rich feed
supplements, ideally a feed used as a supplement should be energy-rich, highly

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 20

digestible, nutrient-dense, economical and without any negative impact on livestock and
the environment.
The choice of Australian farmers was Spirulina platensis, a viable option for a new feed
source for cows.

SPIRULINA IN THE COMPOSITION OF MILK

The fat and protein in the milk is the most important part and this is what payment
systems in Australia are based on. These two components are influenced by the animal's
nutrition and lactation stage. During the grazing season, these elements are lacking so it
is necessary to have a greater intake of energy supplements.
Spirulina algae is a rich source of adequate nutrients and energy and is very useful for
increasing milk production. Kuplys (et al., 2009) observed that in the supplemented
rations with 200 g of Spirulina Platensis the animals produced 6 kg more milk than usual.
This alga can also be used to increase the production of proteins in milk, in fact,
according to other studies, the proteins in milk, with the integration of Spirulina Platensis,
also increase by 1.5 g/Kg.
Thus, although there was a positive response based on this study, the conflicting results
in previous studies indicate the need to further investigate the effects of this seaweed to
understand the relationship between lactation diets.

ARTICLE ON TRIALS IN CATTLE (Carillo et al.2016)

In recent years, microalgae have been used in ruminant diets to enrich meat and milk
on a nutritional level; in fact, ruminants are the right animals to study the effect of
microalgae as they can digest cell wall organisms that are often not processed.
Microalgal supplementation for ruminants is an effective method to reduce saturated
fatty acids and increase concentrations of conjugated linoleic acid and other PUFAs in
ruminant milk (Franklin, et al. 1999, Papadopoulos et al. 2012). These changes in the
fatty acid profile are probably related to changes in the rumen bacterial flora of the
population.
Researchers demonstrated that the addition of microalgae in the diet of dairy cows led
to higher levels of DHA and conjugated linoleic acid in the milk fat and a decrease in
saturated fatty acids. (Franklin, et al. 1999)
Due to the differences in feed types, different diets, different nutrient profiles, based on
the types and levels of supplementation and the length of the experimental period it is
not easy to compare the correlation between the effects of quantitative milk production
and the microalgae feed. (Altomonte, et al. 2018)
It was found that the decrease in microalgae-containing concentrate was balanced by
higher consumption of silage, when the maximum number of microalgae ingested with
no effect on feed was performed in cows, the intake ranged from 4 to 79 g of microalgae/
kg of dry matter in the diet, it is important to note that this reduction effect depends on
the rations supplied together with the microalgae. (Papadopoulos, et al. 2002; Moate, et
al. 2011).
DaSilva, et al. (2016) performed an experiment in which the effects of partial
replacement of dietary ground corn with a microalgae meal (composed of de-oiled
microalgae and soybean shells) in bovine diets were evaluated. It has been observed that
the addition of microalgae in the diet increases digestibility compared to the control diet.

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 21

ARTICLE THAT EXPLAINS THE INCREASE OF CHARACTERS IN MILK WITH THE
ADDITION OF SPIRULINA ALGAE (Manzocchi et al.2020)
The biomass of the cyanobacterium Spirulina is a promising protein source that
represents a valid alternative to soybean meal in the diets of dairy cows. As previously
mentioned, spirulina can be produced on marginal or even landless lands and therefore
does not compete with the land for human food resources.
It is estimated that by 2054, alternative protein sources will account for 33% of total
global protein consumption. Only very few studies have evaluated spirulina as an
ingredient in cow rations.
Studies have reported that the administration of spirulina algae reduces the palatability
of feed concentrates (Moate et al. 2013, Laminen et al. 2019).
It was decided to supplement the diet of grazing cows with 15-30 g/day of spirulina but
this did not affect the BW, BCS, milk production, composition, SCC and blood
antioxidant capacity in the subsequent lactation. (Garcès et al., 2018).
Recently, Till et al. (2019), supplemented the diet of dairy cows with 150 g/day of
another microalga Schizochytrium limacinum was studied, this intake decreased the
proportion of fatty acids in milk and cheese and increased the nutty flavor and the fruity
smell, increasing the astringency and decreasing the creaminess of the cheese.
These results cannot be generalized for all microalgae in dairy cow diets. The effects of
spirulina are different when administered in larger quantities to a diet with lower
percentages of antioxidant compounds than those present in fresh grass, such as hay. In
this respect, the content of phenols in spirulina and their potential effects on the
antioxidant capacity have never been reported in scientific articles or texts.
The benefits of spirulina could include potential improvement in animal health as it is a
good source of micronutrients, agents and antioxidants such as carotenoids, tocopherols
and phenolic acids (Becker, 2013).
The experiment was performed by comparing the blood and milk analyzes of two groups
of cows, the first group with a diet in which spirulina was present, while the second
35 group with a diet in which soy flour was present, therefore a legume flour,
consequently, with a high protein content.
Spirulina contained 20.1% more crude protein than soy. It had a 5 times higher energy
content than soybean meal. Spirulina was rich in tocopherols:

1- α- tocopherol the most abundant;

2- 𝛽 tocopherol;
3- γ-tocopherol;
4- e-tocopherol.

Naturally, vitamin E exists in eight chemical forms (alpha-, beta-, gamma-, and delta-
tocopherol and alpha-, beta-, gamma-, and delta-tocotrienol) that have varying levels of
biological activity. Alpha- (or α-) tocopherol is the only form that is recognized to meet
human requirements.
Furthermore, spirulina contained a significant amount of β-carotene and compound
polyphenols, 90% of which were non-tannic phenols while the remaining 10% were
hydrolysable tannins.
The average body weight of the animals that took part in the experiment was not different
between the various groups and the same happened for milk production as well as

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 22

energy. No effects of the diet change were noted for fat, protein, lactose, urea, and
somatic cell counts. The concentration of β-carotene was higher in total milk and fat of
cows that had spirulina algae added to the diet (4.36 vs 2.59), α-tocopherol and total
phenols and total antioxidant capacity did not differ between the two groups. The milk
of the cows given spirulina algae had a more yellow milk colour than the other group.
Concerning the coagulation time of the rennet, the curd firming rate and the firmness of
the same after 30 minutes from the addition of the rennet were not affected by the change
of diet. Saturated fatty acids (SFA) were higher in spirulina algae-fed cows than in the
other group (118 mg/100g vs 28 mg/100g) however this value was not affected by the
replacement of soybean meal by spirulina in the diet. This substitution resulted in a
reduction of fat in the milk. The proportions of trans-11 were highest in the milk fat of
cows in the first group. Despite the differences observed for some individual MUFAs and
PUFAs, the total proportions of the former were not affected by diet.
The CLA content of the milk was not changed as were the fat indices and α-linolenic
acid. The mean milk fat and protein content of the group that experimented with spirulina
was 4.6 ± 0.15% and 3.7 ± 0.04%, while the other group contained 5.2 ± 0.09% of fat
and 4.1 ± 0.12% of protein.

CHARACTERISTICS OF SPIRULINA AND ITS EFFECTS

Spirulina is much higher in protein than soybean meal, contrary to some previous reports
(Madeira et al., 2017), in the microalga batch, very long chain AF (less than 21 carbon
atoms) could not be detected).
The length of the chain can be explained by the different conditions: temperature, light,
pH, and nutrient availability; the diversity of the strain and the solvent used for the
analysis also depends, on the solubility of the different lipid fractions can also vary.
From all this information we can deduce that the diet was not reduced due to the addition
of 5% spirulina. In this experiment, liquid molasses was added to both experimental
diets, which may have improved the acceptability of the diet and certainly improved the
palatability of the seaweed.
The inclusion of molasses could be a convenient strategy to increase the palatability of
microalgae.

EFFECTS OF SPIRULINA ON THE RUMINAL FERMENTATION

The nutritional composition of spirulina algae has been found to have an impact on
rumen microbes, leading to changes in the rumen fermentation of nutrients. The group
fed with spirulina showed lower levels of volatile fatty acids in the rumen, potentially
due to the protein content and lower proportion of carbohydrates in spirulina compared
to soybean meal. The microbes in the rumen primarily rely on food polysaccharides such
as starch, cellulose, hemicellulose, and pectin to generate energy. However, spirulina
contains only 13% of polysaccharides and a significantly higher percentage of highly-
digestible fibre. Despite the higher carbohydrate intake in the spirulina-supplemented
diet, the reduced rumen fermentation observed may be attributed to incomplete
biohydrogenation of the microalga in the rumen. Corresponding findings by Boeckaert
et al. 2008 support this, where an accumulation of trans-C18:1 in the rumen was found
in animals following a diet with algae supplementation, resulting in changes in the
presence and activity of Butyrivibrio sp. in the rumen.

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 23

Experiment Conclusion: Our findings demonstrate that microalgae can serve as an
effective protein source to replace soy flour in dairy cow diets. Contrary to prior
assumptions, we confirmed that spirulina is palatable when molasses is added, allowing
for easy ingestion by the animals. While there was a reduction in rumen fermentation,
this did not negatively impact milk production or quality. We also observed a decrease
in rumen biohydrogenation and a subsequent lower accumulation in milk. Additionally,
some coagulation properties of the milk were altered, and there was no significant
impact on phenol content, total antioxidant capacity or sensory properties of the milk.
Our experiment revealed that this diet is particularly beneficial for human health due to
the high presence of β-carotene and γ-linolenic acid. To further validate these results,
future research with larger subject groups and longer periods of testing may be necessary
to evaluate the potential effects of bioactive compounds on individual cow and herd
fertility.

INFLUENCE OF CYANOBACTERIA SPIRULINA PLATENSIS ON THE BODY

CONDITION OF LACTATING COWS AND BIOCHEMICAL INDEXES OF MILK (Kulyps
et al. 2009)
The objective of this research was to estimate the influence of the cyanobacteria Spirulina
Platensis on the physical conditions of lactating cows and at the same time to control the
biochemical indices of the milk and finally, if the production was modified.
Two groups of cattle of different Lithuanian breeds in their first lactation period were
considered. The experimental period lasted 90 days, during which the cows were fed
with very similar rations. The first experimental group received a ration with 200 g of
Spirulina cyanobacteria additives per day, mixed with the usually used combined feed.
The body condition of the animals was evaluated according to a 5-point scale system:
dry period, after farrowing, and after the first, second and third month of lactation. Milk
yield and milk quality indices were estimated during the control milking. The experiment
showed that cows fed Spirulina Platensis increased the value4 of milk fat by 8.5-11%,
and each animal produced an average of 34 kg of milk per day, i.e. 6 kg more than the
other group. This method was also cost-effective, as the cost of Spirulina was recouped
by the increased income due to increased production.

MATERIALS AND METHODS

This experimentation was carried out at the Lithuanian Veterinary Academy, for 90 days
between April and June 2007 (end of stable period, beginning of grazing period). As
previously mentioned, data from two groups of Lithuanian cows were analysed,
estimating their body score, milk production and quality. The animals were tethered to
the post, and each group had 2 hours of movement per day, they were fed individually
while water was administered with automatic systems. The cows were milked twice daily
at 5.00 and 16.00 and both groups received 15 kg of silage, 2 kg of hay and 350 g of
combined forage. Each cow of the first experimental group also received 200 g of dry
Spirulina, manually mixed with the combined fodder. The forage ratio corresponded to
the established standards for livestock feed (Jatkauskas et al., 2002). Forage intake was
estimated by first weighing the ingestion and removing any remains from it. As previously
reported, the body condition of the animals was evaluated according to a 5-point scale
(Heinrichs,2004) and was estimated 5 times: in the dry period (30 ± 10 days), after
parturition (1-2 days ), after 1 month of lactation (30 ± 10 days), after 2 months (60 ± 10

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 24

days) and at peak lactation (60 ± 10 days). An evaluation was done after milking and
morning feeding. The examination consisted of the evaluation of different regions: back,
loin and tail.
Milk yield in kg and biochemical indices (fat, protein, lactose, urea concentration and
somatic cell number) were measured in both groups. The milk production was estimated
on the farm while the quality in a state laboratory was according to the standard
methodology.
The standard error and significant differences between the means were established with
the variables using the P test.

RESULTS
Milk yields in the first group, i.e. the experimental one, increased progressively: during
the first month their average milk yield was 4 kg more, in the second month 6 kg more
and in the third month 7 kg more than in the second group i.e. the control group.
Milk production in the control group increased only at the beginning of grazing and
remained the same thereafter. According to the literature (Kupras et al., 2003; Simkus et
al., 2005) "Spirulina platensis" is formed by alkaline elements and other substances which
can modify the reaction of the pre-stomachs. The body condition of the cows in both
groups met the standard; however, after delivery, the experimental group had 15% more
than the control group. At the peak of lactation, the body condition of both groups
decreased, even the animals belonging to the control group did not even match the
standards, in fact, they were almost 10% lower.
With regard to milk composition and biochemical indices, the fat concentration was
quite low in the experimental group during the first and third months of the experiment.
Nutritional factors, the crude fibre of fresh grass and high temperature have had an
impact on the transition period from stables to pastures. By averaging the various checks
in both groups, significantly higher milk production and total milk fat content of the first
group are highlighted.

Figure 17. Fat concentration in milk

As regards the concentration of proteins in the milk in the experimental group, it tended
to increase during the second and third months of the experiment.

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 25

Figure 18. Protein concentration in milk

The lactose concentration increased steadily in both groups, although the first group had
a higher increase.

Figure 19. Lactose concentration in milk

The concentration of urea in the milk indicates the fermentation process in the rumen
and the level of nutrition. If the urea in the milk is low, the ratio is lacking in proteins; if
it is high, the proteins are in excess. In the experiment, the influence that Spirulina has
on the concentration of urea in milk is not estimated.
The number of somatic cells in the milk of both groups of cows tended to vary. In some
cows, from both groups, the number increased but was within the permitted range
throughout the experimental period. According to the data collected, the spirulina algae
had no direct influence.

CONCLUSION
The addition of spirulina algae to the diet during the lactation period positively
influenced milk productivity by 21%.
The composition of the milk and the biochemical indices were similar in both groups of
animals: the fat concentration was rather low in the experimental group during the first
and third months and slightly higher during the second month than also for the protein
part. The lactose concentration increased steadily in both groups; however, in the
experimental group, it was greater than that in the control group.
Spirulina positively influenced bovine oestrus and body condition. In conclusion, it can
be stated that the cost incurred for the use of the microalgae was recovered, even with a
profit.

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 26

10 CONCLUSION
After conducting an extensive investigation into alternative feed sources, with a specific
focus on Spirulina Platensis, it has become evident that further research is required to
fully understand the potential utilization of this algae. From an environmental standpoint,
the utilization of microalgae presents an opportunity to protect and sustainably manage
natural resources, particularly in terms of water and soil conservation. Microalgae have
emerged as a highly promising food resource that has the potential to revolutionize
animal feed, especially with the incorporation of Spirulina Platensis into animal diets.
These microorganisms possess exceptional nutritional qualities, such as proteins,
vitamins, and antioxidants, which remain largely untapped and underutilized. The
inclusion of these compounds in animal diets has been shown to have a positive impact
on their overall health, immune system, productivity, quality, and stability in zooculture.
In summary, the vast potential of microalgae, particularly Spirulina Platensis, warrants
further exploration as it offers a promising natural resource for sustainable animal feed
and supplementary human nutrition.

Given the vast array of microalgae species, undertaking a comprehensive biochemical

analysis is imperative in order to select the most appropriate varieties for utilization in
food technology and as a dietary supplement. Additionally, it is important to consider
the substantial economic commitment required for the complete cycle of microalgae
production, from cultivation to processing and marketing. While algae offer a vegetable-
based product with commendable nutritional value, it remains necessary to conduct
several analyses before their optimal integration into agricultural animal diets can be
assured.

Giuliano Regonesi – Microalgaex Innova4on Center - Istanbul 27

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