Food engineering as a potential solution for mitigating of the detrimental effects of livestock production

Journal of Environmental Treatment Techniques

2019, Volume 7, Issue 2, Pages: 201-210

J. Environ. Treat. Tech.

ISSN: 2309-1185

Journal weblink: http://www.jett.dormaj.com

Food Engineering as a Potential Solution for

Mitigating of the Detrimental Effects of

Livestock Production

Ameneh Bazrafshan¹

and 2

, Tahereh Talaei-Khozani *

PhD Student, Physiology Department, Shiraz University of Medical Sciences, Shiraz Iran

- Maternal and fetal research center, Shiraz University of medical Sciences, Shiraz, Iran

- Anatomy Department, Shiraz University of Medical Sciences, Shiraz Iran

Received: 20/01/2019 Accepted: 30/04/2019 Published: 01/06/2019

Abstract

Global demand for meat is on the rise. Increase in livestock production is the first but not the best solution to supply this

demand. Livestock production leads to an increase in the greenhouse gasses, causing global warming and climate change, which

also has a negative impact on the livestock breeding. Thus, scientists have concentrated on the production of in vitro-engineered

meat which could be tasty, healthy and environmental friendly to substitute livestock meat. In this article, the environmental

impacts of livestock production system on the climate change, water quality and public health are discussed, and then the

artificial meat production technology, its benefits, challenges and consumer’s reactions are reviewed.

Keywords: GHG reduction, Global warming, Food engineering, Laboratory meat production

Introduction¹

concentration flow in the atmosphere leads to global

warming (3).The livestock sector is responsible for 14.5%

of global GHG emissions (10), which may lead to increase

in air and water pollution, land degradation and decrease in

biodiversity (3, 11-13). Generally, livestock sector

contribution in anthropogenic GHG emissions is 53% of

Global demand for livestock products is estimated to

double by 2050 due to increase in human population from

.2 to 9.6 billion by 2050 (1).Global change in lifestyle has

led to increase in demands for agricultural products by

about 70%. Moreover, meat production is expected to

increase from 258 to 455 million tons (2). Livestock

production requires facilities and natural resources for

animal feed production, manure and animal product

processing, transportation and marketing. All of these

contribute to climate change, water and air pollution, land

use change, and other environmental impacts (3). Around

2 4 2

N O, 44% of CH and 5% of CO emission (10). Rise in the

world population and livestock products demand (2) has led

to ideas about other ways of protein production such as new

technologies in producing in vitro engineered meat. It is

also known as a cultured, ‘lab-based’, or artificial meat

(

14). The engineered meat is made of animal stem cells

cultured in a specific medium containing the necessary

nutrients and energy sources for proliferation and

differentiation into muscle cells and adipocytes to produce

a commercial large scale natural tasty meat in the near

future (15-17). Therefore, water, land, nutrients and energy

requirement might be relatively less than livestock

production, because artificial meat is the only muscle tissue

which will be developed without using biological structures

such as respiratory and digestive system. Rapid growth rate

of engineered meat means that preparing it requires shorter

time and also smaller input requirement than that of animal

rearing (18). The engineered meat production has a less

global warming potential and environmental threat than

livestock production (19). Although food preferences,

changes in social habit over time and encouraging the

consumers to use artificial meat instead of the traditional

one is difficult, the engineered meat may become more

8% of the land in the European Union equals to 65% of

the agricultural land, occupied by livestock production

system. Air, water and soil quality, global climate and

biodiversity, biogeochemical cycles of carbon, phosphorus

and nitrogen are affected by livestock production (4).

2 4 2

Carbon dioxide (CO ), methane (CH ), nitrous oxide (N O)

and chlorofluorocarbons are the main greenhouse gases

GHG) involved in the regulation of global temperature (5-

). The normal temperature on earth should be -6°C.GHGs

(

absorb a part of the heat waves from the sun and also its

reflection and then trap them in the atmosphere. GHG

Corresponding

Anatomy department, Shiraz University of Medical

Sciences, Shiraz Iran. Email: talaeit@sums.ac.ir. Phone no:

author:

Tahereh

Talaei-Khozani,

98 7132304372.

Journal of Environmental Treatment Techniques

2019, Volume 7, Issue 2, Pages: 201-210

common as a part of the diet in future. Since current

knowledge of mass production is in its early stages due to

technical, ethical and social issues, it might be assumed that

the production of highly valued meat will face great

technical challenges. Thus, a great range of research will be

required to establish an in vitro engineered meat production

system on a large scale. Current review article discusses the

direct and indirect impacts of livestock production system

on air, water quality and climate change. Here, we will

discuss the necessity to find a proper meat alternative to

reduce livestock products demand; and then we will review

all important aspects of the engineered meat production,

environmental impacts, advantages of engineered meat, and

finally technical and social challenges in the engineered

meat production and market acceptance as a novel food.

emitted from manure by anaerobic decomposition of

organic substance. Globally, anaerobic decomposition of

manure is responsible for4% of the global anthropogenic

methane emissions (3).

1.3 Effect of livestock production system on nitrogen

emission

Livestock contributes to 65% of global anthropogenic

emissions of N

GHG. Livestock produces virtually two thirds of the total

anthropogenic N O emissions (3). Fertilized croplands and

manure are responsible for global increase in N O

O, the most effective of the three major

emissions (25). Current styles confirms that this level will

significantly increase in future (24). Animal wastes, faecal

and urine excretion (26) and manure-induced soil (27) are

responsible for a major amount of the nitrogen emission to

.1 Effect of livestock production system on carbon

the atmosphere. Conversion of nitrogen oxides (NO ) to

dioxide emission(CO

)

nitric acids is taking place in the presence of moisture.

Besides harmful effects of nitric acids to the respiratory

system as well as some materials, it forms acid rain that

return the pollutions to the soil, which can be harmful for

biological ecosystem (3, 4).

Livestock sector is responsible for 9% of carbon

dioxide anthropogenic emissions, when pasture degradation

and deforestation for feedcrop land are taken into

consideration (3). Direct impacts of livestock system in

carbon emission to the atmosphere is lesser than indirect

emissions (3). One of the most important ways of indirect

1.4 Ammonia

emission by livestock is fossil fuel for mineral

Livestock contributes to 64% of global anthropogenic

fertilizer production, which is used in feed production. The

most important routes of GHG emissions from livestock

production system are manure and artificial fertilizers.

Nitrogenous fertilizers, which are used in crop productions,

contribute significantly to GHG emissions. Fossil fuel,

manufacturing process of fertilizer production, packaging,

transport, and application of the fertilizer contribute to emit

emissions of NH , which is mostly from manure (3). Global

anthropogenic ammonia emission was estimated to be

about 58million tons per year in 1993 and it will reached

118million tons per year by 2050 (28). Ammonia and

nitrogen oxides emissions contribute to the formation of

tropospheric ozone (O ), the third most important GHG,

which is direct driver of global warming. The

more than 40 million tons of CO

modern livestock production systems a large amount of

energy is used for diesel machinery involved in seeding,

per year (3). In the

tropospheric ozone induces oxidative stress; hence as a

result, reduces ecosystem productivity, decreases the sink

strength of ecosystems for atmospheric CO , indirectly

leading to global warming (29-31).

herbicides/pesticides

production,

land

preparation,

harvesting, transport, and also a part of energy is used for

electrical devices involved in irriga tion, drying, heating,

cooling, ventilation and etc. (3, 20, 21). Although

1.5 Land use change to feed production for livestock

Since the 1850s,forests and natural fields have been

converted to croplands and pastures for livestock

production (32). Land degradation happens because crop

producers drain the soil resources from nutrient, which

leads to change in physical, chemical, and biological

properties of soil (3). Land use change influences the

natural carbon cycle, because in comparison to the

croplands and pastures the majority of the carbon in soil

and vegetation is sequestered by natural fields such as

forest (33). In addition, land use change can produce other

livestock’s respiration process emits3 billi on tons of CO ,

they are recycled by biological system (3, 22). Transport of

livestock products as well as the livestock products

processing, storage and refrigerated transport require fossil

fuels which are responsible for the CO emission. Part of

the CO emission is produced from shipping the products in

long distances such as, feed delivery to the livestock

production sites, animal products delivery to markets and

raw ingredients delivery around the world (20, 21).

4 2

type of gas emissions like CH and N O by soil

microorganisms that result in global warming (34).

(

.2 Effect of livestock production system on Methane

CH ) emission

Livestock contributes 35–40% of global anthropogenic

emissions. Enteric fermentation may contribute to

per year. Ruminant

animals like cattle, buffaloes, sheep, goats and camels

produce significant amounts of CH during normal digestive

1.6 Effect of livestock production on water pollution and

depletion

emission of 86 million tons of CH

Water has a critical role in the functioning of the

ecosystem, and human activities are the most important

factor in mobilizing this vital natural resource. Freshwater

sources are essential for global sustainability, food

development and main tenance, industrial growth and

ultimately human life (3, 35). However, only 2.5% of total

water resources are fresh water, which has been distributed

unequally. More than 2.3 billion people in 21 countries live

processes (3). Rumen microbial content of animal digestion

system converts the consumed food to digestible feed

during enteric fermentation, which releases a CH by-

product (23). Methane emissions related to livestock will

increase to 60% by 2030, if the livestock production

expansion continued in the same rate (24). Methane is

in water stressed situation (1000 and 1700 m /person/year)

Journal of Environmental Treatment Techniques

2019, Volume 7, Issue 2, Pages: 201-210

like Iran, Yemen, Egypt, Mexico, North China and India (3,

contaminants, including Escherichia Coli (3, 47) and

Salmonella (3, 48) can survive for a long time in the animal

faeces applied as fertilizers on land, leading to water

resources contamination (3). Importantly, many of the

viruses, including Ebola, influenza, Hendra and Nipah are

aresome of the important livestock pathogensthat threat

human health (49). Giardia, Cryptosporidia, Fasciola

hepatica and Fasciola gigantica are also important parasite

infections transm itted through ingestion of contaminated

water or food (3, 50).

6, 37). Water consumption by livestock is considerable,

and with respect to increase in livestock meat production,

livestock water demand has projected to 71% from 1995 to

025 (35) .Around 60% of total water withdrawals by

livestock was from ground water sources in the United

States in 2010 (38). Currently, Iran’s agriculture consumes

about 92% of the freshwater to supply 90% of the food

demands (39). Total water use to produce around 60

million tons of beef every year is higher than the total

freshwater reserves on the planet. For instance, water use

ranges from 11,000 L/kg body weight of beef in Japan to

1.10 Effect of livestock production system on terrestrial

biodiversity

7,800 L/kg in Mexico. This variation in water use is

probably due to differences in local evaporation,

transpiration, livestock production systems, and animal

productivity (40). World population is expected to grow by

around 2.3 billion people between 2009 and 2050,

thusabout two-thirds of the world population would

experience water shortage in the next coming decades (41).

The effect of the livestock sector on water resources are

not well recognized by the decision makers. The total direct

or indirect water usage by the livestock sector is often

overlooked. Livestock production needs service water,

particularly in industrialized farms, to clean the animals and

their units, and also for cooling facilities used for the

animals and their products (milk, meat) (38). Water use in

feed cropland, is much hig her than that of the other water

usage described above (3, 42). Similarly, the influence of

livestock in water depletion is ignored and mainly focused

on water contamination by animal manure and waste.

Intense land usage for livestock production and

rangeland conversion into cropland leads to decrease in

biodiversity. Habitat degradation and change, and land

fragmentation leads to eradicatio n of native species with

invasive non-native plants (51, 52). It is expected that

livestock grazing will lead to a global decrease in main

species abundance in rangeland until 2050 (53). Reduced

species richness via eutrophication, and acidification are

the consequences of nitrogen deposition in soil (54).The

livestock sector has a significant effect on agriculture

syste m and is responsible for 78% of the biodiversity losses

(55, 56).

1.10 How can we reduce livestock impacts onclimate and

water?

Scientists have concentrated on diminishing the GHG

emissions from the livestock sector. A key solution to

reduce GHG emissions could be decrease in meat

consumption. Another way is to shift human dietary style

toward a vegetarian diet or other meat protein alternatives

such as mycoproteins. Most of the people do not like using

vegetable derived meat due to taste, allergic irritation and

psychological issues. Hence, an engineered meat can be an

alternative. Tissue engineering is a new medical technology

to construct a tissue from patient-derived cells seeded onto

scaffolds. Specific biochemical and physical conditions are

provided for cultured cells to produce tissues with

maximum similarity to the original one for transplantation.

We can use the tissue engineering technology to

differentiate muscle cells and adipocytes from farm animal-

derived stem cells by mass production in food industry (14,

18). Summary of the state of the techniques to make tissue

engineered meat, meat production procedure, technical

challenges, benefits, ethical issues and social attitudes are

discussed in the following sections.

.7 Effect of livestock production system on water

pollution

Most of the used water for drinking and servicing in

livestock sector returns to the nature in the form of manure

and wastewater form. Livestock sector contains

significant amount of drug residues, heavy metals,

pathogens and nutrients like nitrogen, phosphorous,

potassium. These substances can cause serious health

hazards in the environment, if they entered into the water

sources or stored in the soil (3, 26).

.8 The main water pollutants related to livestock sector

High concentration of nutrient in the ecosystem leads to

eutrophication, which might be a health threat. Nutrient

ingestion by animals can be high. Most of the ingested

nutrients return to the nature and may become a threat to

water resources (3). Excessive amount of the nutrients in

water resources and eutrophication can lead to the over-

growth of aquatic plants and toxic algae blooms leading to

death of fishes due to oxygen insufficiency, loss of

biodiversity, loss of coral reefs, bad water flavor and odor,

and excessive microbial growth. Livestock-related

activities can significantly accelerate eutrophication, a

usual process in the ageing of lakes, trough high rate of

nutrients and organic substances penetration into the

aquatic ecosystems (43-46).

An overview of the techniques involved in

the in vitro engineered meat

At first, NASA made small quantities of healthy and

safe fish tissues. A testers group smelt the engineered

tissue, but did not eat it to judge how appetizing it was (57).

Then, Dutch researchers showed that the isolated muscle-

derived progenitor cells have long-term expansion and

differentiation capacity (58). In another attempt, electrical

stimulation in skeletal stem cells accelerated sarcomere

assembly in both 2D and 3D conditions. The expanded

stem cells were then differentiated into muscle cells using

chemical/biological clues in the appropriate cell culture

media (59). Finally, Professor Mark from Maastricht

.9 Biological contamination is a health threat-related to

livestock.

Livestock generates many zoonotic micro-organisms

and parasites that threaten human health. Many biological

Journal of Environmental Treatment Techniques

2019, Volume 7, Issue 2, Pages: 201-210

University launched the world's first cultured beef burger

from cow muscle cells and on fifth of August 2013 in a

London press conference, the burger was cooked and eaten

which is a thin tissue strip a mo ng the anchors, and will

form bioartificial muscle (68, 70, 71). Differentiation leads

to satellite cell conversion into primitive muscle cells

containing myotube, which start to express skeletal muscle

proteins like myogenin and muscle myosin heavy chain

(71-73). After differentiation, the muscle will develop

increasing tension between the anchor points. This tension

is the major trigger for protein production. Biochemical

material like growth factors and mechanical nature of the

scaffold are involved in subsequent newly formed myocyte

hypertrophy (70). Passive stretch and tension, and electrical

stimulation induce protein production and force generation.

Electrical stimulation, which has a critical role in muscle

cell differentiation, myoblast maturation and sarcomere

formation, is dependent on coating and matrix stiffness

(74). In addition to contractile proteins, other proteins are

also critical for texture, color and taste of the artificial

muscle tissue. For instance, myoglobin, a heme-carrying

protein, is responsible for the meat pink color, and

determines meat taste as well. It was shown that myoglobin

expression is regulated collectively by activation of

transcription factors including MEF2, NFAT and Sp1,

(

60).

.1 Cell sources and engineered meat production

procedure

The myosatellite cell, a muscle tissue specific stem cell,

and embryonic stem cell which are responsible for muscle

regeneration are used for engineered meat production (15,

7). Adult stem cells have self-renew capacity, with an

unlimited number of cell doublings for tissue regeneration.

Stem cell proliferation and differentiation should be tightly

regulated to avoid uncontrolled cell growth (61).

Proliferation and then differentiation of satellite cells are

the challenging steps to produce meat. The goal of the cell

proliferation phase is to expand the number of cells to be

sufficient for mass production. Current methods in satellite

cells isolation and culturing support about 30 population-

doubling number and 50-70 doubling can probably be

achieved, using proper conditions (62). Collins et al.

showed a major improvement in harvesting satellite cells

using a combination of mild enzymatic digestion and

trituration by keeping them in the replication phase

difference in

intracellular calcium current and low

intracellular oxygen pressure (75). Scaffold removal from

the cell sheet is the main challenge in tissue engineering.

Cell sheets detachment is performed mechanically,

enzymatically and low-tempe rature liftoff from smart

thermoresponsive coatings (76, 77). Generally, 3D-printing

collagen-based meshwork is used as a biocompatible and

biodegradable scaffold. The cells seeded on the scaffolds

are held into a stationary or rotating bioreactor filled with

nutrient. The cells start to fuse and form myotubes, which

is subsequently differentiated into myofibers with the aid of

differentiation media. Soft texture or boneless meat is

produced by using this technique w hich can be used to

make hamburger and sausages (71, 78, 79). In summary,

with the aid of current technology in skeletal muscle

cultivation, making the engineered meat is possible.

(

63).When stem cell niche environment is maintained in the

harvested cells, they can grow more appropriately.

Basement membrane is one of the most important parts of

niche with a regulatory role in proliferation of the stem

cells via signal transduction applied by extracellular matrix

reorganization (61). For example coating the culture dish

with laminin, main basement membrane protein, or

matrigel, could increase the satellite cell proliferation rate

and also myogenic differentiation capacity via Wnt

signaling activation (64). In addition, satellite cell self-

renewal are influence by regulatory circuits such as TGFb1,

Pax7, Notch and Wnt (65). These regulatory mechanisms

can be targeted with specific agonists to induce

proliferation and delay differentiation. Also there are other

non-invasive cell sources for stem cells. The breastmilk

stem cells also have the potential to be differentiated into

different cells derived from mesenchyme (66), including

adipocyte and muscle cells or other cells that can be

potentially used as food such as hepatocytes (67).

Benefits of the engineered meat

In vitro engineered meat is expected to deliver lowered

water usage, GHG emissions, eutrophication, and land use

in comparison to conventional meat production.

Engineered meat is being developed as a healthier and more

efficient alternative to livestock meat. There are several

studies that have investigated the environmental impact of

in vitro engineered meat. Tuom is to et al. used life cycle

assessment to evaluate the environmental impacts of large-

scale engineered meat production. Comparing to

conventional European meat production, the engineered

meat emits nearly 78-96% lesser GHG, 99% lesser land

use, 82-96% lesser water , and 7-45% lesser energy use;

however, the energy consumption depends on the source of

meat; for instance, poultry meat uses lesser energy in

comparison to the engineered meat production (19). On the

contrary, the other comparative study focused on the energy

consumption in supportive industry for engineered meat

production such as culture media production and cleaning

steps. The results indicated that in vitro engineered meat

consumes more industrial energy than livestock meat.

Comparative evaluations of the adverse effect of fabricating

engineered meat on global warming depend on the natural

.2 Mechanical cues

To mimic the natural and 3D structure, a scaffold is

needed with appropriate qualities to allow cell adhesion and

proliferation and tissue recapitulation. Myocytes, as an

anchorage-dependent cell, need enough substrate stiffness

to be functional and contractile. Thus, scaffolds should

provide

a large and flexible surface area to allow

contraction, best medium diffusion and easily detachment

from the culture (68).The best material for scaffold would

be natural and edible like collagen that provides porosity

and flexibility to the structure (69). Protein content and

quality of skeletal muscle tissue is compromised by

expressing contractile proteins in differentiated satellite

cell. The cells are usually seeded in a collagen gel and it is

critical to provide anchoring sites in the culture dish.

Tissue-like matrix is responsible for cell adhesion balance,

contractility, and finally differentiation. Differentiated

satellite cells will organize the collagen gel to the structure,

Journal of Environmental Treatment Techniques

2019, Volume 7, Issue 2, Pages: 201-210

meat sources as well; for instance, the engineered meat

seemed to have a larger global warming potential than pork

or poultry, but less than beef. In vitro meat requires less

land and lower amount of feedstock than livestock (80).

Smetana et al. used different assessment (cradle-to-

plate) to compare engineered meat to a series of meat

alternatives like plant, mycoprotein, and dairy-based, and

delivery are some of the obstacles with large volumes of

cell culture in big bioreactors. Stirring the cell suspension

can be a solution for most of the problems, but the

membrane of the most mammalian cells is not able to

tolerate sheer stress caused by agitation. As a result, the

cells in high scale might suffer from insufficient and

inhomogeneous transfer of oxygen and nutrients due to

limitation in stirring speed (88). Another important problem

chicken, as

conventional meat with the least

environmental impacts. The results revealed that in vitro

engineered meat and mycoprotein-based substitute had the

highest environmental impact, which was due to high

industrial energy requirements for medium cultivation.

Chicken and dairy-based substitute has moderate and soy

meal-based, and insect-based substitute have lowest

environmental impact. The engineered meat just has

beneficial impact on land use and freshwater toxicity. The

overall consequence is that engineered meat production

seems to have lesser environmental impact than some

conventional meat like beef, and probably pork, but more

than chicken and plant-based substitutes (81).

Another benefit is that engineered meat could have less

biological risk and diseases, due to standardized methods in

production. In addition, the composition of engineered meat

could be altered to make the meat healthier or make it for

specialized diet, for example by using higher level of poly-

unsaturated fatty acids in the culture medium. Protein

synthesis by engineered skeletal muscle cells could be

increased by using the optimal biochemical and physical

culture condition (82). One of the main goals of engineered

meat is to slaughter significantly lesser number of animals.

From the perspective of animal activist, this could attract

vegans, vegetarians and others who are interested in

decreasing meat intake due to ethical issues (83). The

engineered meat product could be made in large scale, and

also there is no need for functional integration (84).

in large scale cell culture in big bioreactors is CO

removal.

The CO accumulation affects cell growth which is usually

removed from the culture medium via a combination of

both agitation and air sparging. Due to low rates of used

agitation and sparging, CO accumulation is a limiting stage

in large scale cell culture and growth (89). Even if all the

technical problems of the large scale culture could be

resolved, producing adequate amount of meat for

increasing world population by 2050, would require a huge

number of bioreactors. It was estimated that around one

reactor for ev ery 10 humans is required to supply the meat

demand (84, 85). Huge amount of culture medium also is

needed for this scale of cell culture. Hence, medium

production and storage in large scale will be another

challenge (85). The discarded of the culture medium in a

safe way is also another problem which remains to be

solved. Besides, some of the stem or progenitor cells might

differentiate to undesirable phenotypes. They may modify

epigenetically or undergo karyotypic abnormalities during

culturing. These kinds of undesirable results must be

detected and controlled for human health safety (90, 91).

Several techniques based on physical parameters like

electric conductivity could be used for cell culture process

monitoring, which will be sufficiently strong to guarantee

cell culture quality. For instance Dielectric spectroscopy is

one of the safety control tools, which has been used in the

medical field which might be helpful to monitor the

biological parameters of cell culture (92). In addition, the

nature of commercial cell culture media like the source of

components, extraction method and processing should be

controlled through a life cycle assessment. Considering the

large scale of cell culture for meat production, a range of

innovation through chemical and mechanical engineering

will be required for quality control of cell based products.

Optimization of the cell culture variables, like specific

components of medium and serum, is essential to improve

the efficiency of the culture and consistency of the

products. Feed composition of medium, biochemical and

biophysical condition of culture, and the possible

interactions between medium components should be

defined to produce healthy engineered meat (18).

Technical challenges in the engineered meat

production

Meat tissue engineering has at least three main

challenges including scale, efficiency and taste. To generate

acceptable volume of meat for a large population, cell

culture scale and condition has to be several times higher

than that of used for medical application. Bioreactor design,

selection and production of biomaterial, optimization of

culture medium, tissue conditioning optimization and

quality control of engineered meat such as the genetic

stability of the cells are major problems in scaling up the

engineered meat. The global meat production is around 293

million tons/year (84). There is approximately 5-10 cell/gr

The Good manufacture Practice (GMP) guidance has

been developed to create an awareness of ranging from

important issues in cell and tissue culture. Based on the

GMP guideline, the quality of all used materials, methods

and their application should be confirmed (93).

Optimization of the cell culture condition including both

medium synthesis and serum supplementation is also

important. Serum might be considered as a potential source

of contamination which needs breeding livestock and

slaughtering the animals as well.

of skeletal muscle tissue and, the number of all skeletal

muscle progenitor cells that differentiate into mature

muscle cells and get integrated into skeletal muscle are

around 1.5_10 cells/year (84, 85). This estimation can

provide an assessment for the crude scale of the number of

cells required for industrialization. Currently, capacity of

the large bioreactors for cell suspension culture is

approximately 2 5,000 liters with maximum cell load of 7-

0 cell/ml (84, 86). Providing the surface attachment for

cells in bioreactors is also critical to sustain proliferation

and survival of satellite cells. Different microcarriers are

available to support the large scale of adherent cells in huge

bioreactors (87). Waste washout, oxygen and nutrient

Ultimately, synthetic culture media without using

serum products is the ultimate goal in the cell culture. Since

fetal bovine serum is a supplement in cell culture with

Journal of Environmental Treatment Techniques

2019, Volume 7, Issue 2, Pages: 201-210

unknown composition, which could contain a wide range of

undesirable factors; hence, omitting fetal bovine serum

from cell culture medium seems to be critical. Several types

of culture media have been produced with minimal or no

animal derived components. However, different type of

cells are able to grow in serum-free media, each cell type

needs special medium composition and there is no standard

serum-free media for all cell types (94). Also, several

serum‐free media have been developed for myoblast cell

differentiation with the capability of active tension

generation (95). More studies must be performed to omit

serum from the whole cell culture process to decrease the

dependency on livestock products.

Taste, texture and juiciness of engineered meat are also

a challenge in its production. Meat taste is related to amino

acid and peptide conce ntration and also the intramuscular

fat content of meat (96, 97). Special taste of cooked-meat is

related to the reaction between specific sugars, amino acids

and fatty acids, particularly during heating (98-100). Since

that feed and nutritional conditions of animal as well as

postmortem conditions can affect the taste of the meat due

to protein, sugar and fatty acid oxidation (101, 102),

suggesting the feed conditions of the skeletal muscle

culture could be effective on taste development in

engineered meat. Thus, taste of meat is an important quality

to be investigated in engineered meat production system.

Texture and integrity of meat is determined by the

intramuscular connective tissues, composed of extracellular

molecules like collagens and glycoproteins, and also the

amount and distribution of the adipose tissues. Adipose

tissues development leads to disorganization of the

intramuscular connective tissue structure which causes the

meat tenderness (103). In addition to tenderness, Juiciness

of meat is also related to percentage of fat which varies in

different types of skeletal muscles (104). Medium

composition and feed condition and of cell culture must be

optimized to produce a highly quality engineered meat with

the best taste, texture and juiciness to attract consumers.

repelling the idea of eating engineered meat and moral

issues related to the engineered meat technology and its

application (83). Costumer reactions toward engineered

meat were investigated in three EU countries, Belgium,

Portugal and the United Kingdom. Initial costumer

reactions after learning about engineered meat were

feelings of disgust, fear of the unknown, uncertainly about

safety, health and naturalness. Consumers imagined few

direct personal advantage of engineered meat, but they

accepted possible societal benefits related to environment

and food security (106). A media coverage in 2015 about

artificial meat overemphasized the important role of the

vegetarians in artificial meat acceptance (107). A survey

conducted in the Netherlands showed that only 14% had

heard of artificial meat and claimed to know a little about

that. After explaining the artificial meat technology and its

advantages and disadvantages, about 63% of the people

supported the idea of engineered meat production and 52%

were interested to try it (108). In another study, after giving

customers basic information about engineered meat, only

9% of people rejected, two thirds hesitated and about

quarter supported the idea of trying engineered meat.

Generally, consumers were doubtful about trying the

engineered meat even if it becomes available, those how

are vegetarian will be doubtful about its safety and health

(105). Recently,

a study investigated the effect of

information provision on the attitude toward engineered

meat. Results showed that it can be affected by positive

information about sustainable product (109). Also, it is

important to give the simple and apprehensible information

about final product, but not about production method, to

increase public acceptance (110).

Cost and sensory expectations has appeared as major

obstacles. In 2015, the in vitro-grown burger producer

announced that the burger price from in vitro engineered

meat decreased so that the price will be compatible with a

conventional meat (111). The price drop was astonishing in

just 2 years, which could be a good sign for the engineered

meat commercialization. Taken together, people need

scientific assurance to trust the engineered meat. It can be

claimed that in vitro engineered meat can reduce our stress

on the environment, through reduction in livestock

production, agricultural land and water usage. However, the

possibility of other improvements, like price, quality,

safety, similarity to conventional meat, is difficult to

predict. Consequently, it is crucial to be more transparent

about all aspects of the engineered meat production. A tasty

meat is a basic requirement for societal acceptance, but

beyond that, the societal perception depends on too many

factors. If the detrimental effects of conventional meat

production on environment continually increases and if

tasty engineered meat lead to mitigating the damage to the

nature by preventing animal and plant extinction, this novel

meat may finally become a highly valued food and also as

a regular part of food program (84).

Challenges and outlooks for consumer’s

acceptance of in vitro engineered meat

How will consumers react to engineered meat

production technology? Under which conditions will

customers accept and adapt to eat engineered meat? Will

engineered meat be compatible with highly valued

conventional meat? Customer’s acceptance or rejection

depends on two sets of determinants. The first one is the

personal and societal advantages and health- threatening

risks of the engineered meat. Technology-related issues are

the second set of determinants such as quality control and

safety assurance of cell culture and perceived naturalness of

the engineered meat. Customer cognizant about the

engineered meat production technology is so effective in

acceptance or rejection of the engineered meat by market

(

105). Since, one of the advantages of this technology is

decrease livestock production, animal activists and

vegetarians, who hate the idea of slaughtering animal,

might be conceived to use engineered meat (83). Most

potential objections to the engineered meat were

overviewed by Hopkins et al. including concern about

unknown hazards of the engineered meat technology, doubt

about realness and naturalness of the artificial meat,

Conclusions

Raising world population needs more livestock

products such as meat to supply their food demands. GHG

emissions, water pollution, land use change and

degradation are some of the well-known impacts of

livestock production system. Therefore, finding a proper

Journal of Environmental Treatment Techniques

2019, Volume 7, Issue 2, Pages: 201-210

meat alternative seems to be essential. Engineered meat

could be a good meat substitute with less environmental

impacts. Using the engineered meat could be economically

efficient; if it considers that the consuming budgets for the

prevention of global warming will be decreased

remarkably.

Food and Agriculture Organization of the United

Nations FAO, Rome. 2013.

11.Bellarby J, Tirado R, Leip A, Weiss F, Lesschen JP,

Smith P. Livestock greenhouse gas emissions and

mitigation potential in Europe. Global change biology.

2013;19(1):3-18.

Although the in vitro engineered meat was produced,

cooked and tasted successfully, its production in a large

scale has some technical challenges such as, cell source

selection and providing a biochemical and physical cell

culture condition. Even if the engineered meat becomes

available in the market, it was shown that social acceptance

will be an obstacle which needs more effort to satisfy

consumer to accept and eat the artificial meat.

12.Reynolds C, Crompton L, Mills J. Livestock and

Climate Change Impacts in the Developing World.

Outlook on Agriculture. 2010;39(4):245-8.

13.Thornton PK. Livestock production: recent trends,

future prospects. Philosophical transactions of the Royal

Society of London Series B, Biological sciences.

2010;365(1554):2853-67.

14.Kumar P, Chatli MK, Mehta N, Singh P, Malav OP,

Verma AK. Meat analogues: Health promising

sustainable meat substitutes. Critical reviews in food

science and nutrition. 2017;57(5):923-32.

Acknowledgment

The authors wish to thank research deputy of Shiraz

University of medical Sciences and Mr. H. Argasi at the

Research Consultation Center (RCC) of Shiraz University

of Medical Sciences for his invaluable assistance in editing

this manuscript.

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