“PROANIM / PROBIOTICS IN ANIMAL HUSBANDRY“

(Project № 2020-1-BG01-KA202-079240

USE AND EFFECTS OF PROBIOTICS IN RUMINANTS

Prof.Dr.Kemal ÇELİK1, Prof.Dr.Murat YILMAZ2, Prof.Dr. Harun BAYTEKİN1, Alkan

ÇAĞLI2, Selda MANAV2, Hasan ÇOĞAN2
1. 1.Çanakkale Onsekiz Mart University Faculty of Agriculture, Department of Animal Science
Çanakkale -TURKEY
2.Aydin Adnan Menderes University Faculty of Agriculture, Department of Animal Science,
Aydın -TURKEY

[email protected], [email protected]

Introduction
The ever-increasing world population has increased the demand for animal products worldwide
(FAO, 2015). However, various problems such as poor quality feed, heat stress, global
warming, parasites and diseases, as well as ongoing challenges such as restrictions affect
livestock production (Wanapat et al., 2013; Hernández-Castellano et al., 2019). Ruminants play
an important role in the well-being and livelihood of the global population, making valuable
contributions to the human food chain supply (Wanapat et al., 2015). In the last fifty years,
remarkable progress has been made in the field of livestock, thanks to improvements in health
conditions, genetic selection, nutrition and the use of growth promoters and antibiotics
(Thornton, 2010). The incorporation of nutritional and growth promoters and antibiotic
strategies into commercial animal production has improved livestock health and feed efficiency.
However, antibiotics and growth promoters have caused antibiotic resistance and increased
food-borne allergies, increasing their harmful effects on the environment (Gonzalez-Ronquillo
& Hernandez, 2017; Hao et al., 2014). Although the issue is still under discussion, there is
growing concern among consumers about the human health effects of antibiotics and growth
promoters (Lipsitch et al., 2002). For this purpose, scientists are investigating alternative
strategies to increase the quantity, the quality and the homogeneity of animal products (Musa
et al., 2009; Wanapat et al., 2013).

The poor performance of farm animals has led to an increased need for substitutes. An
alternative on the rise in countries around the world is the probiotic supplement to the diet of
ruminants, either as single or multiple strains. The use of probiotics as an important feed
supplement to optimize production performance has increased worldwide. (Mahesh et al.,
2021).

Ruminant nutritionists have developed many methods of feed supplementation, such as the use
of antibiotic growth promoters, to increase production by limiting the effects of pathogenic
infection on ruminant productivity (Reti et al., 2013; Valero et al., 2014). However, their use
has been significantly reduced on the basis of their health and environmental impact (Gaggia et
al., 2010). In addition, the non-therapeutic use of antibiotics as growth promoters in livestock
diets and for disease prevention was banned by the European Union in 2006 due to the
emergence of resistant pathogenic bacteria and possible contamination of animal products.
Consumers and many countries have implemented the same bans (Russell and Houlihan, 2003;
Jouany and Morgavi, 2007). Therefore, the need to seek more natural feed additives alternatives
to antibiotics stems from consumers' preferences for more natural animal products (Khan et al.,
2016).

The ruminant gastrointestinal tract is home to a wide variety of microbial species that are
directly or indirectly important to the overall well-being of many animals (Stover et al., 2016).
The main fermentative vessel, the rumen, is a complex biological system in which the
degradation, fermentation and conversion of feed materials by microorganisms occur
(McSweeney and Mackie, 2012; Albrao et al., 2014). This fermentative vessel provides an
anaerobic environment (38 to 41 °C constant temperature and 5.5 to 6.9 pH) for microbes
(Dehority, 2003).
Rumen microbes are classified into 3 areas: bacteria, archaea (methanogens), and eukaryotes
(protozoa and fungi). There are over 200 species of rumen bacteria and the population range is
1010 to 1011 per g. Anaerobic fungi in the rumen are divided into 6 genera with populations of
103 to 106 per g and rumen methanogen populations up to 109 per g, while the population range
of bacteriophage and ciliate protozoa is 107 to 109 and 104 to 106 per g, respectively (Choudhury
et al., 2015; Kumar et al., 2009; Wanapat, 2000). The bacterial population is most actively
involved in plant fiber degradation, as evidenced by the fact that bacteria associated with feed
particles make up approximately 50% to 75% of the total microbial population (Minato et al.,
1966). Anaerobic fungi degrade the lignocellulosic components of feed particles. They make
up the smallest proportion (only about 20%) of the rumen microbial biomass (Rezaeian vd.,
2004). Rumen protozoa play an important role in fiber digestion and modulation of fermentation
profiles, by slowing down the production of acids that lower rumen pH and benefiting the rumen
(Arowolo et al., 2018).

Probiotics

FAO/WHO (2002) defines probiotics as “live microorganisms that confer a health benefit on
the host when administered in adequate amounts”. Probiotics are essentially living and non-
pathogenic organisms. Supplements benefit host health by competing with other pathogenic
microbes (Uyeno et al., 2015). Proper balance of rumen microbes is crucial for maintaining
animal health and efficient digestion of feedstuffs (Adjei-Fremah et al., 2018a). Probiotics,
when administered in adequate amounts, can manipulate the rumen microbial ecosystem, feed
digestibility, and degradability in ruminants. However, they increase the bioavailability and
digestive capacity of microbes and lower ruminal pH and lactate levels (Vyas et al., 2014). In
addition, probiotics play a critical role in improving mucosal immunity by activating and
stimulating immune cells, by preventing enteric pathogens from settling in the gut, and by
increasing nutrient utilization and absorption (Chaucheyras-Durand et al., 2012; Hutkins et al.,
2016). Research interest in the application of probiotics is increasing in ruminants, as they help
maintain the balance of the gut microbiota and act as a possible substitute for traditional
antibiotic use. Existing research in animals reveals that probiotics used as feed supplements for
ruminants increase growth and production efficiency and improve health and general well-
being (Bakr et al., 2015; Raabis et al., 2019). Probiotic applications have been proven to
minimize adverse environmental consequences such as methane emissions associated with
ruminant production (Hassan et al., 2020; Jeyanathan et al., 2019).
Probiotics have various properties that are beneficial for animal health and performance (Kassa,
2016; Raabis et al., 2019). They compete with a wide variety of intestinal microflora, survive
in the unfavorable intestinal environment and adhere to the intestinal epithelium
(Bezkorovainy, 2001). In addition, probiotics have certain antimicrobial properties, such as the
ability to improve intestinal health, to increase digestive capacity and to maintain vitality during
use and storage, and they have effective sensory properties and use. Therefore, they can
improve the general immunity of the organism (Collins and Gibson, 1999; Oyetayo and
Oyetayo, 2005; Uyeno et al., 2015).

There are specific variations between different groups of probiotics in relation to their
properties and mechanisms of action. The main LAB (lactic acid producing bacteria) in the
rumen are Streptococcus bovis, Lactobacillus spp. and Enterococci (Kassa, 2016 ). This LAB
(for example, Lactobacilli and Enterococci) continuously supplies lactate in the rumen,
stimulating the growth of lactic acid-utilizing bacterial populations to maintain ruminal pH. An
overpopulation of S. bovis is associated with mild acute or chronic ruminal acidosis (SARA),
and an excess of Escherichia coli has been linked to severe SARA and inflammation (Mazon
vd., 2020; Zapata vd., 2021). Therefore, these LABs have been recommended as direct-feeding
microbes (DFMs) due to their ability to increase rumen microbes that adapt to the presence of
lactate in the rumen and increase LUB (lactic acid-using bacteria) (Elghandour et al., 2015; Seo
et al. However, like Megasphaera elsdenii and Selenomonas ruminantium, LUB stabilizes
ruminal pH by converting lactate to volatile fatty acids (VFAs), while fed a highly fermentable
diet. Therefore, LUB-based DFMs help prevent ruminal acidosis in dairy cows (Aikman et al.,
2011; Mazon et al., 2020). M. elsdenii is one of the major strains of bacteria that utilize lactic
acid in animals fed an easily fermentable diet, as other species of lactate-fermenting bacteria
(S. ruminantium) show catabolite suppression (Chaucheyras-Durand & Durand, 2010). In
addition, M. elsdenii uses maltose, lactate and glucose as energy sources and therefore competes
with LAB for substrate and decreases lactate levels while increasing VFA production (Aikman
et al., 2011; Mazon et al., 2020). Propionibacteria, another potential LUB, ferment lactate to
propionate, thereby increasing propionate production. This, in turn, supports glucose
production, reduces the incidence of postpartum ketosis, or increases energy efficiency by
providing more substrate for lactose synthesis (Stein et al., 2006). In addition, enhanced
propionate can reduce the amount of hydrogen available in the rumen for enteric methane
synthesis (Vyas et al., 2014).
Microorganisms used as probiotics in ruminants

The main and frequently studied microorganisms used as probiotics in ruminant feeds include
bacterial species belonging to the genus Prevotella bryantii , such as Streptococcus , Bacillus ,
Lactobacillus, Enterococcus , Propionibacterium, Bifidobacterium and fungal species including
Saccharomyces and Aspergillus (Gaggìa et al., 2010). Bacteria probiotic strains can be
classified as lactic acid-using bacteria (LUB) and lactic acid-producing bacteria (LAB). The
most prominent probiotic used for commercial purposes in ruminants is Saccharomyces
cerevisiae (Anadón et al., 2006).

Table1. Bacteria that produce lactic acid

Lactobacillus types Lactobacillus brevis
L. lactis
L. reuteri
L. johnsonii
L. fermentum
L. paracasei
L. plantarum
L. acidophilus
Lactobacillus casei ssp. casei
Bifidobacterium types Bifidobacterium longum
B. brev
B. laktis
B. bifidum
B. infantis
B. thermophilum
B. pseudolongum
Enterokok types E. faecalis
E. faecium
Bacteria that use lactic acid
Megasphaera Megasphaera elsdenii
Propionibacterium freudenreichii
P. shermanii
P. freudenreichii
Other Types Bacillus cereus
Saccharomyces cerevisiae
Bacillus licheniformis
Prevotella bryantii
Saccharomyces boulardii
Pediococcus acidilactici
Aspergillus oryzae
Sporolactobacillus inulinus
Candida utilis
(Kulkami vd., 2022)
Several yeast metabolites and yeast cultures are also used as probiotics in ruminants (Shurson,
2018). Yeast culture contains a mixture of yeast biomass and fermentation metabolites
(alcohols, esters, organic acids, etc.) (Mahesh et al., 2021; Shurson, 2018). Meanwhile, dry
brewer's yeast, torula (Candida utilis) and whey yeast are also used in animal nutrition (Mahesh
et al., 2021). Due to their synergistic adhesion effects, combinations of various probiotic strains
can increase positive health effects compared to individual strains (Elghandour et al., 2015).

Live yeast (S. cerevisiae)

Live yeast (S. cerevisiae) is one of the most common and effective probiotics used in the diet
of ruminant animals due to its various functions in stabilizing the rumen environment for
adequate functioning of microbial flora, especially fibrolytic bacteria. Yeast cells are said to be
able to maintain their viability throughout the digestive tract. Yeast (S. cerevisiae) as a feed
additive for ruminants provides organic acids and vitamins to promote the growth of lactic acid
bacteria (LAB) (Khan et al., 2016). It improves rumen metabolism through stability of rumen
pH, increases cellulolytic bacteria population, inhibits bacteria that produce lactate for
substrate, and improves anaerobiosis by clearing oxygen in the rumen (Khan et al., 2016;
Vibhute et al., 2011; Chiquette, 2009; Marden et al., 2008).

The effects of Saccharomyces cerevisiae on the in vitro gas production kinetics and
degradability of the four-fibre feed were investigated at direct addition or pre-incubation. Corn
stove, oat straw, sugar cane pulp and sorghum straw incubated with yeast (S. cerevisiae) at
different doses were given to the animals (in vitro). The direct addition of S. cerevisiae with
corn stove and oat straw or the incubation 72 hours ago improved gas production. Direct
addition of S. cerevisiae to sorghum straw or incubation 72 hours ago increased asymptotic gas
production, gas production rate and initial delay before gas production starts, dry matter and
neutral detergent fiber digestibility. It improved ruminal fermentation of low quality roughage
at 4 to 12 g/kg DM, as gas production is dependent on nutrient availability for rumen
microorganisms (Elgandour et al., 2014).

The effects of Saccharomyces cerevisiae fermentation product on in vitro fermentation and

microbial communities of low quality forages and mixed diets were investigated. S. cerevisiae
fermentation/concentrations were determined as 0, 1, 2, 3 g/L (in vitro). The results showed
that the molar ratio of acetate and propionate increased linearly with the increasing amounts of
S. cerevisiae for RS (rice straw), CS (corn silage), and CSNG (grainless corn silage),
respectively. Microbial protein increased linearly with increasing S. cerevisiae level for RS and
peaked at 1 and 2 g/L S. cerevisiae for CSG (grain corn silage) and CSNG, respectively. The
mushroom population was increased with 1 g/L S. cerevisiae for all roughages except CSNG.
Ruminococcus flavefaciens population increased at 1 or 2 g/L S. cerevisiae for RS, CSNG and
CSG. Total VFA (volatile fatty acid) peaked at 1 g/L in S. cerevisiae for RS, CSNG and CSG
and increased for CS. It was concluded that the addition of S. cerevisiae could improve rumen
fermentation of roughage by stimulating the number of fiber-digesting rumen microbes,
particularly fungal populations (Mao et al., 2014).

The effect of yeast culture (Saccharomyces cerevisiae) on the ruminal microbial population in
buffalo bulls was investigated by (Kumar et al., 2013). Buffaloes were given yeast culture (in
vivo). Compared with the control group, they observed a significant increase in the mean
protozoal count (3.0 × 104 per mL) and the total bacterial count (about 4.0 × 10 per mL).

Bacteria (directly fed microbes)

It has also been stated that they can inhibit the growth of pathogenic organisms, stimulate the
immune system by secreting bacteriocin, and modulate the microbial balance in the
gastrointestinal tract (Khan et al., 2016). Increased milk yield, oil-adjusted milk yield and
increased milk fat content have also been reported with bacterial supplementation (Elghandour
et al., 2015; Khan et al., 2016). Total VFA (volatile fatty acid) and acetate concentrations in the
rumen fluid increased by an average of 19% 0.5, 1, 3, 6 hours after the morning feeding. The
ruminal apparent nutrient digestibility of NDF (neutral detergent fiber), ADF (acid detergent
fiber) and OM (organic matter) was also increased (Qiao et al., 2009).

Effects of rumen dose and ruminal fermentation on cellulolytic populations in the recurrence
of (NJ) Ruminococcus flavefaciens isolated from a probiotic mixture in feed- or concentrate-
fed dairy cows were researched by Chiquette et al. (2007). They observed that during repeated
dosing, NJ altered the abundance of other cellulolytic bacterial populations in the rumen
compared to periods without dosing. NJ has also improved the digestibility of Timothy hay
with a highly concentrated diet. The increase in persistence of NJ was also noticed in cows at
weeks of dosing.

The effect of microbial supplementation of L. plantarum, Pediococcusacidilactici,

Enterococcus faecium and L. lactis to grass silage on the fermentation properties of dairy cows
was investigated. The ration was prepared as L. plantarum, Pediococcus acidilactici,
Enterococcus faecium and L. lactis / 5×105 colony-forming units/g fresh herb mixture (in vivo).
A significant increase in the lactic acid concentration was observed and it was noted that the
acetic acid concentration in the silage was significantly reduced. When the animals were fed,
the rumen germ count of the cows fed with silage was 13.9% higher than the control. The ratio
of acetic acid to propionic acid in the rumen of cows treated with silage was found to be 1.19%
lower . While the protein nitrogen and total nitrogen content of the rumen was 5.17 mg/100 mL
and 3.37 mg/100 mL higher compared to the control, rumen protein synthesis was improved
(Jatkauskas and Vrotniakien, 2007).

The effect of probiotics on growth and development in ruminants

Probiotics increase microbial ecology, microbial protein synthesis and nutrient absorption while
maintaining a more desirable ruminal pH and lactate concentration to support better weight gain
in ruminants (Bayatkouhsar et al., 2013; Direkvandi et al., 2020b; Elaref et al., 2020; Musa et
al., 2009; Pudasaini and Dhital, 2017).

In newborn calves, the normal population of beneficial microbes such as Bifidobacteria and
Lactobacillus is minimal, but researchers have observed increased growth rate after probiotic
supplementation of these microbes (Stein et al., 2006). As an alternative to antibiotics,
probiotics have been reported to increase the body weight gain of ruminants by increasing
efficiency of nutrient utilization, increasing nitrogen retention, and reducing nutrient excretion.
It was observed that immune adequacy and weight gain of young calves increased with the
supplementation of Lactobacillus acidophilus and Lactobacillus plantarum (Al-Saiady, 2010).

Diets fortified with Pediococcus pentosaceus and Pediococcus acidilactici and fed to lambs
after weaning increased mean daily gain (+25.2g/lamb), final body weight (+3.16kg), FCR (-
1, 18) and overall gain (+ 2.11 kg) (Saleem et al., 2017). Similarly, the average daily weight
gain (ADG) of Sohagi lambs fed with dry yeast improved by 18.4%, thereby increasing overall
growth performance (Elaref et al., 2020).

Kowalski et al. (2009) reported greater ADG and final body weight gain in cows fed Bacillus
licheniformis and Bacillus subtilis. In another study, Sahu et al. (2019) reported that newborn
calves fed cultures of Streptococcus faecium and Lactobacillus plantarum for up to 5 days had
70% less diarrhea with improved feed conversion efficiency.

Hillal et al. (2011) observed a 7.2% increase in ADG for growing lambs fed a mixture of yeast
and probiotic bacteria (i.e., S. cerevisiae, Lactobacillus, Aspergillus, and Streptococcus).
Additionally, some researchers have reported that supplementation of multi-species bacteria
and yeast (Lactobacillus acidophilus, Saccharomyces cerevisiae, Enterococcus faecium,
Aspergillus oryzae, active dry yeast culture) increased overall body weight gain and ADG in
dairy cows and calves (Adjei-Fremah et al. ., 2016 , 2018b). For example, Direkvandi et al.
(2020a, b) reported improvement in feed efficiency, ADG, N uptake, absorption and retention,
and DM digestibility of Arabian lambs fed with multistrain DFM (Lactobacillus fermentum and
L. plantarum) and yeast (S. cerevisiae and M. elsdenii plus Lactobacilli). The tendency for
multi-species probiotic combinations to improve growth performance was due to the
improvement in fiber-degrading bacterial populations and microbial protein synthesis.

Probiotic use and increase in milk production in ruminants

Song et al. (2012) stated that probiotics are included in animal nutrition to improve animal
health and also to ensure food safety. In addition, it is said that yeast provides more nutrient
bioavailability by optimizing rumen function and as a result improves the milk production
performance and provides digestive comfort of the animal (Maamouri et al., 2014; Ayad et al.,
2013).

Maamouri et al. (2014) observed an increase of 1.1 kg/day in milk yield production of Tunisian
Holstein Friesian cows, which was approximately 8% higher than the non-fortified group.
Researchers also reported a higher yield of milk fat (53 g/cow) and protein (41.7 g/cow) in
yeast-supplemented cows compared to 47 and 38.7 g/cow per day in the control group,
respectively. In addition, Moallem et al. (2008) used live yeast (1 g/4 kg) and found that the
average daily milk yield of dairy cows increased by 4.1%.

It has been reported that differences in the response of dairy cows to yeast probiotics among
various studies may be a result of the types of diets offered, the types and doses of yeast used,
and the physiological and nutritional status of the animals (Williams et al., 1991). In addition,
supplementation of S. cerevisiae (5 g/cow) increased milk production by 22.5% in 100 days,
by 13.1% in 200 days and by 14.9% at 305 days (Majdoub-Mathlouthi and Kraiem, 2009).
Likewise, Ayad et al., (2013) reported that supplementation with yeast probiotic (S. cerevisiae)
improved milk production at 42 days in cows by 23%, and the lactation peak of cows lasted 1
week longer than control cows.

Xu et al. (2017) reported a 37% increase in milk yield of dairy cows fed in a tropical
environment and fed supplemental Lactobacillus plantarum P-8 and Lactobacillus casei.
Increased milk yield in dairy animals supplemented with probiotics in India has been attributed
to a higher absorption rate of microbial-derived amino acids and a reduced incidence of mastitis
(Alhussien & Dang, 2018). Milk yield in tropical Tunisian Holstein Friesian cows in mid-
lactation increased by approximately 1.1 kg/day, which was higher than the control group
(Maamouri et al., 2014). These authors also noted higher milk fat (53 g/cow/day) and protein
(41.7 g/cow/day) yields in yeast-supplemented cows.

During the transition period, in their second or third lactation, Murrah buffaloes supplemented
with a fermented yeast culture (Saccharomyces cerevisiae, 24 g/day) increased milk yield and
milk fat content by 13% and 7.5%, respectively (Ahmad Para et al., 2019). Most of the
beneficial effects of probiotics on milk yield and composition are attributed to changes in
volatile fatty acids in the rumen, as well as the indirect effect of probiotics on the number of
cellulolytic and fiber-degrading bacteria (Adjei-Fremah et al., 2018b).

Use of probiotics in ruminants and their effect on methane

Methane production from ruminants has been identified as the largest and only source of
anthropogenic CH4 (Mathison et al., 1998). Animals release methane as part of their natural
digestive processes. The rumen is home to billions of microbes, including bacteria,
methanogens, protozoa, and fungi. These microbes feed themselves to produce volatile fatty
acids (VFAs), carbon dioxide, ammonia and methane. While VFAs are used by animals as an
energy source, gases are removed in a variety of ways. Estimated CH4 emissions from enteric
fermentation are 17-30% of global production (Igbal et al., 2008).

According to Johnson and Johnson (1995), cattle can produce 250-500 liters of methane per
animal per day. Cattle typically lose 2-15% of the energy they ingest as raw methane (Giger-
Reverdin and Sauvant 2000). Therefore, there are two important benefits of reducing methane
losses from cattle. First, less methane means a lower concentration of greenhouse gas (GHG)
in the atmosphere. Second, less methane means increased productivity of livestock production
and increased income for farmers.
 Figure1. Schematic diagram of methane production (Sun et al., 2021)

Probiotics are microbial feed additives that directly affect rumen fermentation and result in
increased animal productivity. The most commonly used probiotics are yeast and Aspergillus
oryzae. Some products available guarantee a high number of live yeast cells and are sold as live
yeast, while other products are sold as yeast cultures containing both the yeast cells and the
medium on which they are grown. Yeast cultures are assumed to reduce methane production in
four ways:

1) by increasing the production of butyrate or propionate (Lila et al., 2004),

2) by reducing protozoan numbers (Newbold et al., 1998),

3) by promoting acetogenesis (Chaucheyras et al., 1995) and

4) by improving animal productivity (Bruno et al., 2005).

In a review of in vitro studies, Wallace and Newbold (1993) found that probiotics increase
productivity by 7-8%, resulting in reduced methane production per unit product (milk or meat)
in dairy cows and growing cattle.

Mwenya et al., (2004) reported that sheep produced significantly less methane when 0.4% yeast
culture was included in the basal hay and 30% concentrate diet. However, McGinn et al. (2004)
reported that the reduction in methane was small and not statistically significant. However, it
can be argued that short-term incubations are not suitable for studying the effects of yeast on
rumen fermentation.

The effects of S. cerevisiae are controlled by an effect on the number and activity of microbes
in the rumen, and it may take longer than 24-48 hours to fully understand the effects of yeast.
Rumen-simulating fermenters provide a convenient tool to study the long-term effects of
additives on rumen fermentation and can be used to screen for the effects of S. cerevisiae strains
in the rumen (Newbold, 2003). Some of the studies have shown that the methane suppression
effects of probiotics are not consistent (Newbold et al., 1995a, b). There is a need to determine
the diet and management situation where probiotics can yield consistent results.

Reducing methane by changing feed type, quality and concentration of ruminants

Studies were conducted to determine the effect of starch-based and fiber-based concentrates on
enteric CH4 production. Lovett et al. (2005) showed that increased use of fiber-based feed on
pasture reduced enteric methane per kilogram of animal product. Positive responses in methane
reduction of high starch-based feed (grains) have been reported by others (Beauchemin and
McGinn 2005). Increasing starch in the diet changes rumen volatile fatty acid (VFAs)
concentrations to form less acetate and more propionate and limits the hydrogen requirement
for methanogenesis. In addition, as the propionate ratio increases, the pH decreases, and this
reduces the methanogenic activity (Walichnowski and Lawrence 1982). In addition,
concentrated feeding has been shown to reduce methane output by reducing the protozoal
population (Van Soest 1982). However, increased concentration levels can cause health
problems such as acidosis (Igbal et al., 2008).

Benchaar et al. (2001) used a modeling approach to evaluate the effectiveness of different
available feeding strategies to reduce methane production from ruminants. The contents of the
simulated strategy are the dry matter intake (DMI), the roughage/concentrate ratio, the nature
of the concentrate (fiber concentrate vs starchy concentrate), the starch type (slowly degraded
or rapidly degraded), the feed types (legume and grass), the feed maturity, the roughage feed
preservation method (dried and siloed), the feed processing and improvement and the
supplementation of low quality roughage (straw). It was noted that methane production was
reduced by increasing the DMI and the concentrate ratio in the diet (-7% and -40%).

The use of more digestible feed (less mature and processed feed) resulted in a reduction in
methane production (-15% and -21%). Methane production in legumes was found to be lower
than grass feed (-28%). Legumes generally have a higher dry matter intake. This has reduced
methane emissions per unit milk or meat production. Hegarty (1999b) found that methane
production per gram of live weight gain was significantly reduced when animals were
transferred from low digestible pastures to highly digestible pastures. Methane emissions per
unit of feed intake can be reduced by using selected grass varieties to improve animal
performance.

However, when we look at the literature, it has not been seen that applications for the production
of high quality roughage have been tested under field conditions. Confidence in the accuracy
of methane measurement technology is essential to implement abatement procedures under
field conditions. While many methods have been used to measure or calculate methane
emissions from ruminants (eg Flux gradient techniques, Boundary layer techniques), the
preferred method for grazing animals is a sulfur hexafluoride (SF6) tracer technique developed
by Johnson et al. (1994a).

Other methods have a very high error of detecting small differences that are important in
inventory, regulatory or animal science research. In addition, pasture improvement will be a
valuable option when compared with the reduction of animal numbers. Although the methane
cost of production (methane/kg beef or milk) will decrease due to the increase in feed intake
associated with the improved digestibility of the pasture, daily (or annual) methane emissions
by the individual will either increase or remain the same (Igbal et al., 2008).

Results

The administration of probiotics to ruminants has yielded significant results by increasing their
productivity and performance, as well as improving their health status. The inclusion of
probiotics in the diet of farm animals can not only improve health status, but also ensure food
security. It has been shown by the presence of higher functional components that supplementing
ruminants with probiotics increases milk production, milk quality and milk composition.
Celluloid bacteria and VFAs from healthy rumen fermentation play a key role in meeting MP
production as well as 70-80% of the total energy requirements of mature ruminant animals.
Between 15% and 70% of the diets given to ruminant animals consist of cellulose and
hemicellulose. The ability of probiotics to improve food digestibility and feed intake in
ruminant animals can be attributed to the proliferation and growth of ruminal celluloid bacteria
and prevention of rumen acidosis. Yeast supplementation of the lactating cows increases feed
intake frequency, fiber degradation and digestibility of nutrients in the rumen.

During weaning, calves become highly susceptible to pathogen colonization in their gut as a
result of an abrupt change in immature gut microbiota and feed composition. The rapid
adaptation to the change in feed composition, the microorganisms living in the gastrointestinal
tract (GIT) of calves and the rapid colonization of beneficial microorganisms are of great
importance in the maturation and development of ruminants. Besides the effect of colostrum
(breast milk) in ruminants, the inclusion of probiotics in the immature GITs of young ruminants
can improve health.

Ruminate animal husbandry in the world is becoming increasingly intensive. Especially in

ruminants fed with mixed and processed feeds and ration contents that are not in their nature,
methane gas production naturally increases. It is not ruminant animals that are responsible for
methane gas production, but industrialization and sectorial incentives that make them feed
incorrectly or against their nature. Methane release will be much less in animals that are fed
with roughage or graze in nature without receiving concentrated feed. By reducing methane
release in ruminants, by improving animal feeding methods such as natural pasture feeding, by
increasing roughage ratios, or by taking measures such as the use of probiotics, both the animal
can be fed more economically and less methane production and a lower concentration of
greenhouse gas (GHG) in the atmosphere is formed. . Less methane production by ruminant
animals will mean increasing productivity of livestock production and increasing income for
farmers.

As a result, in today's conditions, many feed additives such as probiotics can be used to ensure
good ruminal fermentation in ruminant animal breeding. These products will help provide a
better rumen ecosystem, prevent excessive degradation of rumen soluble protein, preserve
rumen ammonia and reduce energy losses. The use of probiotics will increase the use of protein
and energy in the rumen, as well as positively affect the milk production and herd health of
dairy cattle.
References:

Abrão, F. O., Duarte, E. R., Freitas, C. E. S., Vieira, E. A., Geraseev, L. C., da Silva-Hughes,
A. F., ... & Rodrigues, N. M. (2014). Characterization of fungi from ruminal fluid of beef cattle
with different ages and raised in tropical lignified pastures. Current microbiology, 69(5), 649-
659.
Adjei-Fremah, S., Ekwemalor, K., Asiamah, E. K., Ismail, H., Ibrahim, S., & Worku, M.
(2018a). Effect of probiotic supplementation on growth and global gene expression in dairy
cows. Journal of Applied Animal Research, 46(1), 257–263
Adjei-Fremah, S., Ekwemalor, K., Worku, M., & Ibrahim, S. (2018b). Probiotics and Ruminant
Health. Probiotics - Current Knowledge and Future Prospects.
Adjei-Fremah, S., Ekwemalor, K., Worku, M., & Ibrahim, S. (2018b). Probiotics and Ruminant
Health. Probiotics - Current Knowledge and Future Prospects.
Ahmad Para, I., Singh, M., Punetha, M., Hussain Dar, A., Ahmad Naik, M., Salam Teli, A., &
Gupta, D. (2019). Milk production and feed efficiencies as affected by dietary yeast
(Saccharomyces cerevisiae) supplementation during the transition period in Murrah buffaloes.
Biological Rhythm Research, 50(5), 718–725
Aikman, P. C., Henning, P. H., Humphries, D. J., & Horn, C. H. (2011). Rumen pH and
fermentation characteristics in dairy cows supplemented with Megasphaera elsdenii NCIMB
41125 in early lactation. Journal of Dairy Science, 94(6), 2840–2849
Alhussien, M. N., & Dang, A. K. (2018). Milk somatic cells, factors influencing their release,
future prospects, and practical utility in dairy animals: An overview. Veterinary World, 11(5),
562–577
Al-Saiady.M. Y. (2010). Effect of probiotic bacteria on immunoglobulin G concentration and
other blood components of newborn calves. Journal of Animal and Veterinary Advances, 9,
504–609. In Journal of Animal and Veterinary Advances (Vol. 9, Issue 3, pp. 604–609)
Anadón, A., Martínez-Larrañaga, M. R., & Aranzazu Martínez, M. (2006). Probiotics for
animal nutrition in the European Union. Regulation and safety assessment. Regulatory
Toxicology and Pharmacology: RTP, 45(1), 91–95
Arowolo, M. A., & He, J. (2018). Use of probiotics and botanical extracts to improve ruminant
production in the tropics: A review. Animal Nutrition, 4(3), 241-249.
Bakr, H. A., Hassan, M. S., Giadinis, N. D., Panousis, N., Ostojic-Andric, D., El-Tawab, A., &
Bojkovski, J. (2015). Effect of Saccharomyces cerevisiae supplementation on health and
performance of dairy cows during transition and early lactation period. Biotechnology in
Animal Husbandry, 31(3), 349–364
Bayatkouhsar, J., Tahmasebi, A. M., Naserian, A. A., Mokarram, R. R., & Valizadeh, R. (2013).
Effects of supplementation of lactic acid bacteria on growth performance, blood metabolites
and fecal coliform and lactobacilli of young dairy calves. Animal Feed Science and
Technology, 186(1–2), 1–11
Beauchemin KA, McGinn SM (2005) Methane emissions from feedlot cattle fed barley or corn
diets. J Anim Sci 83:653–661
Benchaar C, Pomar C, Chiquette J (2001) Evaluation of diet strategies to reduce methane
production in ruminants: a modeling approach. Can J Anim Sci 81:563–574
Bezkorovainy, A. (2001). Probiotics: determinants of survival and growth in the gut. The
American Journal of Clinical Nutrition, 73(2 Suppl), 399S-405S.
Bruno RGS, Rutigliano HM, Cerri RLA, Robinson PH, Santos JEP (2005) Effect of feeding a
Saccharomyces Cerevisiae yeast culture on lactation performance of dairy cows under heat
stress in ruminant nutrition, feed additives and Feedstuffs. J Anim Sci 85: Proquest Biology
Journals, 310 pp
Chaucheyras F, Fonty G, Bertin G, Gouet P (1995) In-vitro H2 utilisation by a ruminal
acetogenic bacterium cultivated alone or in association with an Archea methanogen is
stimulated by a probiotic strain of Saccharomyces cerevisiae. Appl Environ Microbiol
61:3466–3467
Chaucheyras-Durand, F., & Durand, H. (2010). Probiotics in animal nutrition and health.
Beneficial Microbes, 1(1), 3–9
Chaucheyras-Durand, Frederique, Chevaux, E., Martin, C., & Forano, E. (2012). Use of Yeast
Probiotics in Ruminants: Effects and Mechanisms of Action on Rumen pH, Fibre Degradation,
and Microbiota According to the Diet. Probiotic in Animals, October 2012
Choudhury, P., Salem, A., Jena, R., Kumar, S., Singh, R., & Puniya, A. (2015). Rumen
Microbiology: An Overview. Rumen Microbiology: From Evolution To Revolution, 3-16. doi:
10.1007/978-81-322-2401-3_1
Collins, M. D., & Gibson, G. R. (1999). Probiotics, prebiotics, and synbiotics: approaches for
modulating the microbial ecology of the gut. The American Journal of Clinical Nutrition, 69(5),
1052S-1057S
D. Song, S. Ibrahim, S. Hayek, Recent application of probiotics in food and agricultural science.
Probiotics, INTECH (2012), pp. 3-36
Dehority, B. A. (2003). Rumen microbiology (Vol. 372). Nottingham: Nottingham University
Press.
Direkvandi, E., Mohammadabadi, T., & Salem, A. Z. M. (2020a). Oral administration of lactate
producing bacteria alone or combined with Saccharomyces cerevisiae and Megasphaera
elsdenii on performance of fattening lambs. Journal of Applied Animal Research, 48(1), 235–
243
Direkvandi, E., Mohammadabadi, T., & Salem, A. Z. M. (2020b). Effect of microbial feed
additives on growth performance, microbial protein synthesis, and rumen microbial population
in growing lambs. Translational Animal Science, 4(4), txaa203
Elaref, M. Y., Hamdon, H. A. M., Nayel, U. A., Salem, A. Z. M., & Anele, U. Y. (2020).
Influence of dietary supplementation of yeast on milk composition and lactation curve behavior
of Sohagi ewes, and the growth performance of their newborn lambs. Small Ruminant
Research, 191, 106176
Elghandour, M. M. Y., Salem, A. Z. M., Castañeda, J. S. M., Camacho, L. M., Kholif, A. E., &
Chagoyán, J. C. V. (2015). Direct-fed microbes: A tool for improving the utilization of low
quality roughages in ruminants. Journal of Integrative Agriculture, 14(3), 526–533
Elghandour, M. M. Y., Salem, A. Z. M., Castañeda, J. S. M., Camacho, L. M., Kholif, A. E.,
& Chagoyán, J. C. V. (2015). Direct-fed microbes: A tool for improving the utilization of low
quality roughages in ruminants. Journal of Integrative Agriculture, 14(3), 526–533
F. Gaggia, P. Mattarelli, B. Biavati Probiotics and prebiotics in animal feeding for safe food
production Int J Food Microbiol, 141 (2010), pp. S15-S28
FAO/WHO. Guidelines for the Evaluation of Probiotics 531 in Food. Report of a Joint
FAO/WHO Working Group on Drafting Guidelines for the Evaluation of Probiotics in Food.
2002.
G.H. Qiao, A.S. Shan, N. Ma, Q.Q. Ma, Z.W. Sun, Effect of supplemental bacillus cultures on
rumen fermentation and milk yield in Chinese Holstein cows, J Anim Physiol Anim Nutr, 94
(2009), pp. 429-436
Gaggìa, F., Mattarelli, P., & Biavati, B. (2010). Probiotics and prebiotics in animal feeding for
safe food production. International Journal of Food Microbiology, 141 Suppl, S15-28
Giger-Reverdin S, Sauvant D (2000) Methane production in sheep in relation to concentrate
feed composition from bibliographic data. In: Ledin I, Morand-Fehr P (eds) 8th Seminar of the
sub-network on nutrition of the FAO-CIHEAM inter-regional cooperative research and
development network on sheep and goats. INRA, Cahiers-Options-Mediterraneennes, Grignon,
pp 43–46
Gonzalez-Ronquillo, M., & Hernandez, J. C. (2017). Antibiotic and synthetic growth promoters
in animal diets: Review of impact and analytical methods. Food Control, 72, 255–267
H.L. Mao, Hua-long Mao, J.K. Wang, J.X. Liu, I. Yoon, Effects of Saccharomyces cerevisiae
fermentation product on in vitro fermentation and microbial communities of low-quality
forages and mixed diets, J Anim Sci, 91 (2014), pp. 3291-3298
Hao, H., Cheng, G., Iqbal, Z., Ai, X., Hussain, H. I., Huang, L., Dai, M., Wang, Y., Liu, Z., &
Yuan, Z. (2014). Benefits and risks of antimicrobial use in food-producing animals. Frontiers
in Microbiology, 5, 288
Hassan, A., Gado, H., Anele, U. Y., Berasain, M. A. M., & Salem, A. Z. M. (2020). Influence
of dietary probiotic inclusion on growth performance, nutrient utilization, ruminal fermentation
activities and methane production in growing lambs. Animal Biotechnology, 31(4), 365–372
Hegarty RS (1999a) Reducing rumen methane emissions through elimination of rumen
protozoa. Aust J Agric Res 50:1321–1328.
Hernández-Castellano, L. E., Nally, J. E., Lindahl, J., Wanapat, M., Alhidary, I. A., Fangueiro,
D., Grace, D., Ratto, M., Bambou, J. C., & de Almeida, A. M. (2019). Dairy science and health
in the tropics: challenges and opportunities for the next decades. Tropical Animal Health and
Production, 51(5), 1009–1017
Hillal, H., El-Sayaad, G., & Abdella, M. (2011). Effect of growth promoters (probiotics)
supplementation on performance, rumen activity and some blood constituents in growing
lambs. Archives Animal Breeding, 54(6), 607–617
Hutkins, R. W., Krumbeck, J. A., Bindels, L. B., Cani, P. D., Fahey, G. J., Goh, Y. J., Hamaker,
B., Martens, E. C., Mills, D. A., Rastal, R. A., Vaughan, E., & Sanders, M. E. (2016). Prebiotics:
why definitions matter. Current Opinion in Biotechnology, 37, 1–7
Iqbal, M. F., Cheng, Y. F., Zhu, W. Y., & Zeshan, B. (2008). Mitigation of ruminant methane
production: current strategies, constraints and future options. World Journal of Microbiology
and Biotechnology, 24(12), 2747-2755.
J. Chiquette, G. Talbot, F. Markwell, N. Nili, R.J. Forster, Repeated ruminal dosing of
Ruminococcus flavefaciens NJ along with a probiotic mixture in forage or concentrate-fed dairy
cows: effect on ruminal fermentation, cellulolytic populations and in sacco digestibility, Can J
Anim Sci, 87 (2007), pp. 237-249
J. Chiquette, The role of probiotics in promoting dairy production, WCDS Adv Dairy Technol,
21 (2009), pp. 143-157
J. Jatkauskas, V. Vrotniakien, Effect of L-plantarum, Pediococcusacidilactici, Enterococcus
faecium and L-lactis microbial supplementation of grass silage on the fermentation
characteristics in the rumen of dairy cows, Vet Zootec, 40 (2007), pp. 29-34
J.B. Russell, A.J. Houlihan, Ionophoreresistance of ruminal bacteria and its potential impact on
human health, FEMS Microbiol Rev, 27 (2003), pp. 65-74
J.P. Jouany, D.P. Morgavi Use of ‘natural’ products as alternatives to antibiotic feed additives
in ruminant production Animal, 10 (2007), pp. 1443-1466
J.P. Marden, C. Julien, V. Monteils, E. Auclair, R. Moncoulon, C. Bayourthe, How does live
yeast differ from sodium bicarbonate to stabilize ruminal pH in high yielding dairy cows?, J
Dairy Sci, 91 (2008), pp. 3528-3535
Jeyanathan, J., Martin, C., Eugène, M., Ferlay, A., Popova, M., & Morgavi, D. P. (2019).
Bacterial direct-fed microbials fail to reduce methane emissions in primiparous lactating dairy
cows. Journal of Animal Science and Biotechnology, 10(1), 41
Johnson K, Huyler M, Westberg H, Lamb B, Zimmerman P (1994a) Measurement of methane
emissions from ruminant livestock using an SF6 tracer technique. Environ Sci Technol 28:359–
362.
Johnson KA, Johnson DE (1995) Methane emission from cattle. J Anim Sci 73:2483–2492
K.L. Reti, M.C. Thomas, L.J. Yanke, L.B. Selinger, G.D. Inglis, Effect of antimicrobial growth
promoter administration on the intestinal microbiota of beef cattle, Gut Pathog, 5 (8) (2013),
pp. 1-16
Kassa, S. R. (2016). Role of probiotics in rumen fermentation and animal performance: A
review. International Journal of Livestock Production, 7(5), 24–32
Kowalski, Z. M., Górka, P., Schlagheck, A., Jagusiak, W., Micek, P., & Strzetelski, J. (2009).
Performance of Holstein calves fed milk-replacer and starter mixture supplemented with
probiotic feed additive. Journal of Animal and Feed Sciences, 18(3), 399–411.
Kulkarni, N. A., Chethan, H. S., Srivastava, R., & Gabbur, A. B. (2022). Role of probiotics in
ruminant nutrition as natural modulators of health and productivity of animals in tropical
countries: an overview. Tropical Animal Health and Production, 54(2), 1-15.
Kumar, S., Puniya, A., Puniya, M., Dagar, S., Sirohi, S., Singh, K., & Griffith, G. (2009).
Factors affecting rumen methanogens and methane mitigation strategies. World Journal Of
Microbiology And Biotechnology, 25(9), 1557-1566. doi: 10.1007/s11274-009-0041-3
L. Majdoub-Mathlouthi, K.L. Kraiem, Effects of feeding Saccharomyces cerevisiae Sc 47 to
dairy cows on milk yield and milk components, in Tunisian conditions, Livest Res Rural Dev,
21 (2009), pp. 271-284
Lila ZA, Mohammed N, Yasui T, Kurokawa Y, Kanda S, Itabashi H (2004) Effects of a twin
strain of Saccharomyces cerevisiae live cells on mixed ruminal microorganism fermentation in
vitro. J Anim Sci 82:1847–1854
Lipsitch, M., Singer, R. S., & Levin, B. R. (2002). Antibiotics in agriculture: when is it time to
close the barn door? Proceedings of the National Academy of Sciences of the United States of
America, 99(9), 5752–5754
Lovett DK, Stack LJ, Lovell S, Callan J, Flynn B, Hawkins M et al (2005) Manipulating enteric
methane emissions and animal performance of late-lactation dairy cows through concentrate
supplementation at pasture. J Dairy Sci 88:2836–2842
M. Kumar, V. Kumar, D. Roy, R. Kushwaha, S. Vaiswani, Application of herbal feed additives
in animal nutrition-a review, Int J Lives Res., 4 (2014), pp. 1-8
M.A. Ayad, B. Benallou, M.S. Saim, M.A. Samadi, T. Meziane, Impact of feeding yeast culture
on milk yield, milk components, and blood components in Algerian dairy herds, J Vet Sci
Technol, 4 (2013), pp. 135-140
M.M.Y. Elghandour, A.Z.M. Salem, J.S.M. Castaneda, L.M. Camacho, A.E. Kholif, J.C.V.
Chagoya, Direct-fed microbes: a tool for improving the utilization of low-quality roughages in
ruminants, J Integr Agric, 14 (2015), pp. 526-533
M.V. Valero, R.M. Prado, F. Zawadzki, C.E. Eiras, G.S. Madrona, I.N. Prado Propolis and
essential oils additives in the diets improved animal performance and feed efficiency of bulls
finished in feedlot Acta Scientiarum Anim Sci, 36 (2014), pp. 419-426
Mahesh, M. S., Mohanta, R. K., & Patra, A. K. (2021). Probiotics in Livestock and Poultry
Nutrition and Health. In G. Goel & A. Kumar (Eds.), Advances in Probiotics for Sustainable
Food and Medicine (pp. 149–179). Springer Singapore.
Mathison GW, Okine EK, McAllister TA, Dong Y, Galbraith J, Dmytruk OIN (1998) Reducing
methane emissions from ruminant animals. J Appl Anim Res 14:1–28
Mazon, G., Campler, M. R., Holcomb, C., Bewley, J. M., & Costa, J. H. C. (2020). Effects of
a Megasphaera elsdenii oral drench on reticulorumen pH dynamics in lactating dairy cows
under subacute ruminal acidosis challenge. Animal Feed Science and Technology, 261, 114404
McSweeney, C., & Mackie, R. (2012). Micro-organisms and ruminant digestion: state of
knowledge, trends and future prospects. Background study paper, 61.
Mınato, H., Endo, A., Hıguchı, M., Ootomo, Y., & Uemura, T. (1966). Ecologıcal Treatıse On
The Rumen Fermentatıon. The Journal Of General And Applied Microbiology, 12(1), 39-52.
doi: 10.2323/jgam.12.39
Musa, H. H., Wu, S. L., Zhu, C. H., Seri, H. I., & Zhu, G. Q. (2009). The potential benefits of
probiotics in animal production and health. Journal of Animal and Veterinary Advances, 8(2),
313–321
Musa, H. H., Wu, S. L., Zhu, C. H., Seri, H. I., & Zhu, G. Q. (2009). The potential benefits of
probiotics in animal production and health. Journal of Animal and Veterinary Advances, 8(2),
313–321.
Mwenya B, Santosa Sar C, Gamo Y, Kobayashi T, Takahashi J (2004) Effects of including beta
1-4 galacto-oligosaccharides, lactic acid bacteria or yeast culture on methanogenesis as well as
energy and nitrogen metabolism in sheep. Anim Feed Sci Technol 115:313–326.
Newbold CI, McIntosh FM, Wallace RJ (1998) Changes in the microbial population of a rumen
simulating fermenter in response to yeast culture. Can J Anim Sci 78:241–244
Newbold CJ (2003) Probiotics. Principles for use in ruminant nutrition. In: van Vuuren AM
(ed) The role of probiotics in animal nutrition and their link to the demands of European
consumers. TNO Publications, The Netherlands
Newbold CJ, Lassalas B, Jouany JP (1995a) The importance of methanogenesis associated with
ciliate protozoa in ruminal methane production in vitro. Lett Appl Microbiol 21:230–234.
Newbold CJ, Wallace RJ, Chen XB, McIntosh FM (1995b) Different strains of Saccharomyces
cerevisiae differ in their effects on ruminal bacterial numbers in-vitro and in sheep. J Anim Sci
73:1811–1818
O. Maamouri, H. Selmi, N. M'Hamd, Effects of yeast (Saccharomyces cerevisiae) feed
supplement on milk production and its composition in Tunisian Holstein Friesian cows, Sci
Agric Bohem, 45 (2014), pp. 170-174
Oyetayo, V.O., and Oyetayo, (2005). Potential of probiotics as biotherapeutic agents targeting
the innate immune system. African Journal of Biotechnology, 4(2), 123–127.
P.E.V. Williams, C.A.G. Tait, G.M. Innes, C.J. Newbold, Effects of the inclusion of yeast
culture (Saccharomyces cerevisiae plus growth medium) in the diet of dairy cows on milk yield
and forage degradation and fermentation patterns in the rumen of steers, J Anim Sci, 69 (1991),
pp. 3016-3026
Pudasaini, R., & Dhital, B. (2017). Effect of Supplementing Rice Bran and Wheat Bran with
Probiotics on Growth Performance of Khari Kids. International Journal of Applied Sciences
and Biotechnology, 5(4), 430–433
R.U. Khan, N. Shabana, D. Kuldeep, K. Karthik, T. Ruchi, M.A. Mutassim, et al. Direct-fed
microbial: beneficial applications, modes of action and prospects as a safe tool for enhancing
ruminant production and safeguarding health, Int J Pharm, 12 (2016), pp. 220-231
Raabis, S., Li, W., & Cersosimo, L. (2019). Effects and immune responses of probiotic
treatment in ruminants. Veterinary Immunology and Immunopathology, 208, 58–66.
Raabis, S., Li, W., & Cersosimo, L. (2019). Effects and immune responses of probiotic
treatment in ruminants. Veterinary Immunology and Immunopathology, 208, 58–66.
Rezaeian, M., Beakes, G., & Parker, D. (2004). Distribution and estimation of anaerobic
zoosporic fungi along the digestive tracts of sheep. Mycological Research, 108(10), 1227-1233.
doi: 10.1017/s0953756204000929
Sahu, J., Yadav, A., Kumari, T., & Pal, P. (2019). Probiotic supplementation to produce
healthier calves : A short note. The Pharma Innovation Journal, 8(3), 494–495.
Seo, J. K., Kim, S., Kim, M. H., Upadhaya, S. D., Kam, D. K., & Ha, J. K. (2010). Direct-fed
microbials for ruminant animals. Asian-Australasian Journal of Animal Sciences, 23(12),
1657–1667.
Shurson, G. C. (2018). Yeast and yeast derivatives in feed additives and ingredients: Sources,
characteristics, animal responses, and quantification methods. Animal Feed Science and
Technology, 235(November 2017), 60–76
Stein, D. R., Allen, D. T., Perry, E. B., Bruner, J. C., Gates, K. W., Rehberger, T. G., Mertz,
K., Jones, D., & Spicer, L. J. (2006). Effects of feeding propionibacteria to dairy cows on milk
yield, milk components, and reproduction. Journal of Dairy Science, 89(1), 111–125
Stein, D. R., Allen, D. T., Perry, E. B., Bruner, J. C., Gates, K. W., Rehberger, T. G., Mertz,
K., Jones, D., & Spicer, L. J. (2006). Effects of feeding propionibacteria to dairy cows on milk
yield, milk components, and reproduction. Journal of Dairy Science, 89(1), 111–125
Stover MG, Watson RR, Collier R. Pre- and probiotic supplementation in ruminant livestock
production. 2016. p. 25e36.
Sun, K., Liu, H., Fan, H., Liu, T., & Zheng, C. (2021). Research progress on the application of
feed additives in ruminal methane emission reduction: a review. PeerJ, 9, e11151.
Thornton, P. K. (2010). Livestock production: recent trends, future prospects. Philosophical
Transactions of the Royal Society of London. Series B, Biological Sciences, 365(1554), 2853–
2867
U. Moallem, H. Lehrer, L. Livshitz, M. Zachut, S. Yakoby, The effects of live yeast
supplementation to dairy cows during the hot season on production, feed efficiency, and
digestibility, J Dairy Sci, 92 (2008), pp. 343-351
Uyeno, Y., Shigemori, S., & Shimosato, T. (2015). Effect of probiotics/prebiotics on cattle
health and productivity. Microbes and Environments, 30(2), 126–132.
Uyeno, Y., Shigemori, S., & Shimosato, T. (2015). Effect of probiotics/prebiotics on cattle
health and productivity. Microbes and Environments, 30(2), 126–132.
V.M. Vibhute, R.R. Shelke, S.D. Chavan, S.P. Nage, Effect of probiotics supplementation on
the performance of lactating crossbred cows, Vet World, 4 (2011), pp. 557-561
Van Soest PJ (1982) Nutritional ecology of the ruminant. O & B Books Inc, Corvallis
Vyas, D., Uwizeye, A., Mohammed, R., Yang, W. Z., Walker, N. D., & Beauchemin, K. A.
(2014). The effects of active dried and killed dried yeast on subacute ruminal acidosis, ruminal
fermentation, and nutrient digestibility in beef heifers. Journal of Animal Science, 92(2), 724–
732
Vyas, D., Uwizeye, A., Mohammed, R., Yang, W. Z., Walker, N. D., & Beauchemin, K. A.
(2014). The effects of active dried and killed dried yeast on subacute ruminal acidosis, ruminal
fermentation, and nutrient digestibility in beef heifers. Journal of Animal Science, 92(2), 724–
732
Walichnowski Z, Lawrence SG (1982) Studies into the effects of cadmium and low pH upon
methane production. Hydrobiologia 91–92:1573–5117
Wallace RJ, Newbold CJ (1993) Rumen fermentation and its manipulation: the development of
yeast cultures as feed additives. In: Lyons TP (ed) Biotechnology in the feed industry. Alltech
Technical Publications, Nicholasville, Kentucky, pp 173–192
Wanapat, M. (1990). Nutritional aspects of ruminant production in Southeast Asia with special
reference to Thailand. Khon Kaen University, Faculty of Agriculture, Department of Animal
Science.
Wanapat, M., & Kang, S. (2013). World buffalo production: Challenges in meat and milk
production, and mitigation of methane emission. Buffalo Bulletin, 32(SPEC. ISSUE 1), 1–21.
Wanapat, M., Cherdthong, A., Phesatcha, K., & Kang, S. (2015). Dietary sources and their
effects on animal production and environmental sustainability. Animal Nutrition, 1(3), 96–103
Wanapat, M., Kang, S., & Polyorach, S. (2013). Development of feeding systems and strategies
of supplementation to enhance rumen fermentation and ruminant production in the tropics.
Journal of Animal Science and Biotechnology, 4(1),
Zapata, O., Cervantes, A., Barreras, A., Monge-Navarro, F., González-Vizcarra, V. M.,
Estrada-Angulo, A., Urías-Estrada, J. D., Corona, L., Zinn, R. A., Martínez-Alvarez, I. G., &
Plascencia, A. (2021). Effects of single or combined supplementation of probiotics and
prebiotics on ruminal fermentation, ruminal bacteria and total tract digestion in lambs. Small
Ruminant Research, 204, 106538
Zhu, W., Zhang, B. X., Yao, K. Y., Yoon, I., Chung, Y. H., Wang, J. K., & Liu, J. X. (2016).
Effects of Supplemental Levels of Saccharomyces cerevisiae Fermentation Product on
Lactation Performance in Dairy Cows under Heat Stress. Asian-Australasian Journal of Animal
Sciences, 29(6), 801–806