ВИКОРИСТАННЯ МІКРОБНИХ ДОБАВОК ПІД ЧАС ПРИГОТУВАННЯ СИЛОСУ З ЛЮЦЕРНИ (огляд літератури)
Alfalfa poses challenges for ensiling because of its elevated protein levels, low amounts of water-soluble carbohydrates, low dry matter content, and high buffering capacity. As a result, there has been a recent push to improve silage production using additives. In recent years, silage additives hav...
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| author | Бешараті, M. Лакнер, M. |
| author_facet | Бешараті, M. Лакнер, M. |
| author_institution_txt_mv | [
{
"author": "M. Бешараті",
"institution": "Тебрізський університет, Іран"
},
{
"author": "M. Лакнер",
"institution": "Університет прикладних наук «Технікум Відня», Австрія"
}
] |
| author_sort | Бешараті, M. |
| baseUrl_str | https://smic.in.ua/index.php/journal/oai |
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| datestamp_date | 2026-07-22T10:10:52Z |
| description | Alfalfa poses challenges for ensiling because of its elevated protein levels, low amounts of water-soluble carbohydrates, low dry matter content, and high buffering capacity. As a result, there has been a recent push to improve silage production using additives. In recent years, silage additives have been employed to enhance the quality of alfalfa silage. Bacterial additives are employed to enhance the quality of crop silage, with a particular emphasis on hay silage. A primary objective of incorporating lactic acid bacteria into silage is to inhibit the proliferation of undesirable microorganisms, including Clostridium and Enterobacteriaceae. This is achieved by swiftly elevating the hydrogen ion concentration to a threshold that is inhospitable for the growth of these detrimental bacteria. Recent insights into the functions of bacterial additives in crop silage suggest significant potential for enhancing silage, not just as a fermented feed, but also to deliver probiotic substances that can benefit animal health. This article provides a comprehensive overview of the silage preparation process and critically assesses a range of studies concerning the quality of silage, as well as the impact of bacterial additives on alfalfa silage. The quality of silage can be enhanced by incorporating different bacterial inoculants, which help during fermentation, storage, and feeding by improving fermentation processes, encouraging beneficial microbial diversity, and inhibiting harmful microorganisms. Alfalfa is the most important forage, and microbial additives can enhance its silage preparation in a cost-effective and environmentally friendly way. |
| doi_str_mv | 10.35868/1997-3004.40.3-36 |
| first_indexed | 2025-09-17T09:33:22Z |
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3
МІКРОБІОЛОГІЯ КОРМІВ
Сільськогосподарська мікробіологія. 2024. Вип. 40. С. 3–36.
ISSN 1997-3004
https://doi.org/10.35868/1997-3004.40.3-36
UDC 579.64:636.085
USE OF MICROBIAL ADDITIVES IN ALFALFA
SILAGE PREPARATION
(a review)
M. Besharati1, M. Lackner2
1University of Tabriz
29 Bahman Boulevard, Tabriz, East Azerbaijan Province, Iran
2University of Applied Sciences Technikum Wien
6 Hoechstaedtplatz, Vienna, 1200, Austria; e-mail: maximilian.lackner@technikum-wien.at
Alfalfa poses challenges for ensiling because of its elevated protein levels, low amounts of wa-
ter-soluble carbohydrates, low dry matter content, and high buffering capacity. As a result, there
has been a recent push to improve silage production using additives. In recent years, silage addi-
tives have been employed to enhance the quality of alfalfa silage. Bacterial additives are employed
to enhance the quality of crop silage, with a particular emphasis on hay silage. A primary objective
of incorporating lactic acid bacteria into silage is to inhibit the proliferation of undesirable micro-
organisms, including Clostridium and Enterobacteriaceae. This is achieved by swiftly elevating the
hydrogen ion concentration to a threshold that is inhospitable for the growth of these detrimental
bacteria. Recent insights into the functions of bacterial additives in crop silage suggest significant
potential for enhancing silage, not just as a fermented feed, but also to deliver probiotic substances
that can benefit animal health. This article provides a comprehensive overview of the silage prepa-
ration process and critically assesses a range of studies concerning the quality of silage, as well as
the impact of bacterial additives on alfalfa silage. The quality of silage can be enhanced by incor-
porating different bacterial inoculants, which help during fermentation, storage, and feeding by im-
proving fermentation processes, encouraging beneficial microbial diversity, and inhibiting harmful
microorganisms. Alfalfa is the most important forage, and microbial additives can enhance its si-
lage preparation in a cost-effective and environmentally friendly way.
Key words: alfalfa, bacterial additive, buffer capacity, silage, wilting.
Introduction. High-quality silage is achie-
ved by reducing the activity of plant enzymes
and harmful microorganisms while promoting
the growth of lactic acid bacteria. Alfalfa (Medi-
cago sativa) is the most widely grown forage
globally, known for its exceptional nutritional
value, high yield potential, and other beneficial
characteristics, earning it the title of “queen of
forage”. For several decades, alfalfa has been
highly significant, not just as a nutritious fodder
crop for dairy farming, but also for its positive
role in various health and environmental issues
(Fig. 1) [73].
Typically, alfalfa is stored as dry fodder af-
ter harvesting, but in recent years, the produc-
tion of alfalfa silage has gained popularity
among ranchers. This shift is attributed to sev-
eral factors: minimizing the loss of leaves and
nutrients in the field post-harvest, reducing sto-
rage delays caused by adverse weather, and the
fact that silage is more compatible with mecha-
nization in industrial livestock operations [22;
33]. Various attributes of forage, including the
types of silage plant species, dry matter levels,
water-soluble carbohydrates, buffering capacity
(which indicates resistance to pH fluctuations),
and the interplay of these elements on the mi-
crobes found in the fodder, influence the out-
come of the fermentation process. Alfalfa, in
particular, tends to lose a significant portion of
© M. Besharati, M. Lackner, 2024
4
its nutritional value during the ensiling process
because of its low levels of soluble carbohy-
drates and its hollow stems (Fig. 2) [33; 135].
Fig. 1. Uses of Alfalfa as a multipurpose
crop [73].
Fig. 2. Optimum level of quality parame-
ters for good silage [62].
DMC = dry mass content; NDF = neutral
detergent fiber; ADF = acid detergent
fiber;
CF = crude fiber; IVDMD = In Vitro Dry
Matter Digestibility;
WPY = Water-Soluble Carbohydrates.
Concerning general optimum levels for
good silage (Fig. 2), the following can be stated:
– Dry Mass Content (DMC): Typically, the
ideal DMC for good silage ranges from 30 % to
35 %. Silage that is too wet can lead to poor
fermentation, while silage that is too dry can be
difficult to compact and ferment properly.
– Neutral Detergent Fiber (NDF): Optimal
levels for NDF in silage should be between
40 % to 60 % of dry matter. NDF measures the
total fiber content, and it affects the intake po-
tential of the silage by animals.
– Acid Detergent Fiber (ADF): ADF levels
should ideally range from 25 % to 35 %. Lower
ADF values are generally preferred as they are
associated with higher digestibility.
– Crude Fiber (CF): Although crude fiber is
less commonly used as a primary parameter, it
generally aligns with ADF and NDF levels, and
lower values indicate better quality.
– In Vitro Dry Matter Digestibility
(IVDMD): High IVDMD values indicate better
silage quality, with optimal ranges typically be-
ing above 65 %. This parameter measures the
digestibility of the dry matter content.
– Water-Soluble Carbohydrates (WSC): For
good silage, the WSC content should be high
(above 8–10 % of dry matter), as these sugars
are essential for efficient fermentation.
– pH: Silage should have a pH between 3.8
to 4.2, which ensures proper fermentation and
preservation.
– Ammonia Nitrogen (% of total N): Sho-
uld ideally be less than 10 %. Higher levels in-
dicate poor fermentation.
For several decades, the addition of carbo-
hydrate sources and bacterial additives have
been studied to improve the quality of silage.
Citrus pomace, tomato pomace, apple pomace,
sugar beet pomace, and residues from pistachio
peeling are among the by-products of the trans-
formation and agricultural industries, which are
introduced as a potential source for animal feed
[135; 140]. These materials are produced in the
respective factories depending on the season of
fruit production and are mostly thrown away
without use, which causes environmental pollu-
tion when being mismanaged, such as CH4 re-
lease, and disposal costs. Since adding carbohy-
drate sources can enhance fermentation but does
not stop proteolysis — caused by heterolactic
fermentation and a gradual drop in pH [3] — the
combined use of microbial additives and an ap-
propriate carbohydrate source can lead to im-
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 40.
5
proved fermentation and silage with greater nu-
tritional value. Wilting is used to enhance the
dry matter content of forage prior to ensiling.
Silage that has a very low dry matter content is
typically linked to higher effluent production
and clostridium fermentation, whereas silage
with a high dry matter content does not pack ef-
ficiently and significantly decreases aerobic sta-
bility. The ideal dry matter level for ensiling al-
falfa varies based on the type of silo design, as
well as environmental and management factors
[23; 126].
The low critical pH that inhibits the growth
of Сlostridium is directly influenced by the
moisture levels in the plant. In most cases, when
ensiling materials have high moisture content,
except for those with significant soluble carbo-
hydrate levels, Сlostridium fermentation occurs,
resulting in silage that is of poor quality and nu-
tritional value. Increased humidity leads to a re-
duction in the optimal intake of dry matter and
generates a substantial amount of wastewater,
which is highly nutritious and was found to be
200 times more environmentally harmful than
human sewage [36; 83; 138]. Wilting enhances
the lactate to acetate ratio by promoting a more
uniform and restricted fermentation, thereby
boosting the nutritional quality of silage. Even
in the absence of additives, wilting can lead to
improved silage quality by optimizing the fer-
mentation process [142]. In the United States
and Europe, bacterial additives are frequently
utilized for preserving silage. These products
consist of uniform strains that generate lactic ac-
id, including Lactobacillus plantarum, Entero-
coccus faecium, and Pedicococcus spp. The use
of these additives leads to a quicker reduction in
pH, a lower final pH, an increased ratio of lac-
tate to acetate, and reduced levels of ethanol and
ammonia nitrogen, ultimately resulting in a 1 %
to 2 % enhancement in the recovery of nutrients
from silage [50].
Commercially available heterogeneous ad-
ditives, including Lactobacillus buccaneri and
bacteria that produce propionic acid, generate
additional volatile fatty acids. This process
helps inhibit fungal growth and safeguards si-
lages that are prone to spoilage in aerobic envi-
ronments [50; 78]. The effectiveness of bacteria
in silage is primarily influenced by the moisture
level of the silage and is restricted by high dry
matter content. Since the nutritional quality of
ensiled legumes is significantly affected by the
degree of protein breakdown during the ensiling
process, there has been a recent focus on en-
hancing the quality of fodder to create feed with
optimal nutritional value for livestock. Proper
management of alfalfa silage is crucial for pro-
ducing high-quality silage [103].
History of silage preparation
Silage is a material produced through the
controlled fermentation of moist agricultural
products. The process of making silage and the
location where it occurs are both referred to as
silage (Fig. 3) [70].
Fig. 3. Silage preparation in a traditional
trench silo (Source: https://www.fao.org/4/
x6512e/X6512E05.htm).
The modern techniques for producing fod-
der silage were developed by the French farmer
Goffart, who released his initial book on corn
silage preparation in 1877. Additionally, art-
work from ancient Egypt, dating back to 1000 to
1500 years before Christ, indicates that the
Egyptians had knowledge of silage preparation
for the preservation of agricultural goods. In the
Mediterranean area, airtight storage of fodder
has played a crucial role in the preservation of
agricultural products [47; 89].
Suitable plants for making silage
Silage can be produced from a variety of
plants (Fig. 4). Some plants are specifically
grown for silage production, while others are
used for silage due to an excess supply. Ideal
characteristics for plants intended for silage in-
clude an appropriate level of fermentable sub-
stances, particularly water-soluble carbohy-
drates. Additionally, the materials should have
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 40.
6
Fig. 4. Sources of silages production from various crops [116].
a relatively low buffering capacity and contain
more than 20 % dry matter. The physical struc-
ture of the plants should also allow for easy
compaction into silage after harvesting [21; 25;
89]. The quality and nutritional content of silage
fodder is influenced by various biological and
technical factors, such as the type and species of
the agricultural product, its maturity stage and
dry matter content at harvest, the size of the
chopped fodder pieces, the rate of silage dis-
charge, the weather conditions during harvest, and
the use of additives, among others [104; 136].
The amount of fiber carbohydrates
The primary goal of producing silage is to
preserve the nutritional quality of the core agri-
cultural product as effectively as possible. This
preservation is accomplished through acidifica-
tion and the establishment of an oxygen-free
environment, which serves as the primary sub-
strate for the bacterial fermentation of water-
soluble carbohydrates. Bacteria convert fermen-
table carbohydrates into organic acids, particu-
larly lactic acid and acetic acid [23; 70; 136].
Carbohydrates found in plants are divided
into two main types: structural carbohydrates
and non-structural carbohydrates. Structural
carbohydrates encompass hemicellulose, pectin,
and fibrous polysaccharides like cellulose [38].
In grass (Gramineae, Poaceae) forage, hemicel-
lulose features a primary xylan chain made up
of D-xylose units, along with side chains that
include methylglucuronic acid, as well as some
glucose, galactose, and arabinose. Pectin is
made up of branched chains of D-galacturonic
acid units linked by (1-4) α bonds. Fibrous poly-
saccharides consist of cellulose, which is a line-
ar polymer of D-glucose units. The sugars pre-
sent in these structural carbohydrates are not
readily available to lactic acid bacteria for fer-
mentation. However, they can become accessi-
ble through hydrolysis by plant enzymes or en-
zymes added during the silage preparation pro-
cess [27; 32]. Legumes have lower levels of wa-
ter-soluble carbohydrates than grasses and con-
tain minimal dry matter. The primary polysac-
charide found in temperate grasses is fructan,
whereas legumes primarily contain starch. Starch
does not dissolve in cold water and is not classi-
fied as a water-soluble carbohydrate, making it
unsuitable for most lactic acid bacteria [23; 25].
Forage protein amount
In plant growth, around 75 % to 95 % of ni-
trogen is found as true protein. In silage, this
percentage may drop to between 30 % and
50 %. The primary cause of this issue is not mi-
crobial activity; rather, the activity of protease
enzymes is significant in this context. Plant pro-
teases serve various functions and different spe-
cies exhibit varying stability based on pH and
temperature. Legume forages like clover and al-
falfa contain high levels of degradable crude
protein. In contrast, byproducts from cereals,
such as wheat straw, rice straw, and corn straw,
are low in crude protein [23; 53].
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 40.
7
Factors affecting the breakdown of pro-
teins
Dry Matter
When plants wilt, the pH remains relatively
stable, so any reduction in protein breakdown is
linked to the rise in dry matter. Slight wilting of
materials can actually enhance protein degrada-
tion by preventing acidification [18; 23; 119].
pH reduction
If the pH drops gradually, the breakdown of
proteins will rise. Plant protease enzymes func-
tion effectively at pH levels above 4 [18; 23].
This contradicts the assertion that plants do not
possess active protease enzymes at pH levels
below 4 [83]. Current knowledge indicates that
numerous plant proteases have optimal pH le-
vels that are lower than this threshold. The ac-
tivity of these enzymes declines steadily bet-
ween pH 4 and 6 [81; 82].
Temperature
Since plant protease enzymes function best
at elevated temperatures, therefore raising the
temperature in silage enhances their activity
[18; 23; 82].
Plant Species
The influence of species on the rate of pro-
tein breakdown remains uncertain. The extent of
protein degradation while materials are stored in
the silo is influenced by the proteolytic potential
and the rate of pH decline. Proteolytic potential
refers to the overall activity of the protease en-
zyme, as well as the availability and affinity of
the materials. This trait varies among different
species and is likely influenced by crop ma-
nagement practices and environmental condi-
tions [18; 23; 82].
Time
The activity of proteolysis diminishes over
time during the process of silage preparation. In
the case of both corn and alfalfa, proteolytic ac-
tivity takes place within the first hour of the ini-
tial day following silage preparation, and it de-
clines after five days. This reduction in proteo-
lysis over time is not influenced by the dry mat-
ter content of alfalfa [18].
Natural process of forage fermentation
and silage production
Once the plant is harvested from the field,
its cells remain active, and the microorganisms
in the silo generate CO2 and heat. The process
of turning fresh fodder into preferred silage in-
volves several fermentation stages, which take
approximately 21 days to complete (Fig. 5)
[12; 60].
Respiration stage of the plant
The respiration process in plant cells and
microorganisms within the silo generates water,
carbon dioxide, and heat. This phase is referred
Fig. 5. The ensiling process [5].
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 40.
8
to as aerobic respiration since it requires oxy-
gen. As carbon dioxide levels rise, the rate of
cellular respiration diminishes and ultimately
ceases. The heat generated by aerobic bacteria
raises the temperature of the silage. Typically,
this respiration phase lasts between 3 to 5 hours,
depending on the oxygen availability in the silo
[12; 60].
Acetic acid production stage
This phase starts when the silage runs out of
oxygen, leading to the growth of anaerobic bac-
teria. Bacteria that produce acetic acid utilize
the carbohydrates in the silage, transforming
them into acetic acid, which initiates the acidifi-
cation process and lowers the pH from 6 to 5.
The reduction in pH results in a decline in the
population of acetic acid-producing bacteria.
A swift drop in pH restricts the function of en-
zymes that break down proteins. This fermenta-
tion stage typically lasts between one to two
days [12; 60; 70].
Lactic acid production stage
This phase starts when the quantity of ace-
tic acid-producing bacteria from the earlier pha-
se declines. Lowering the pH of the silage envi-
ronment enhances the growth of lactic acid-pro-
ducing bacteria, leading to the decomposition of
carbohydrates and the formation of lactic acid,
ethanol, mannitol, and carbon dioxide [12; 60].
Peak stage of lactic acid production
This is the longest phase of fermentation,
and is the third stage. Lactic acid production be-
gins in this stage, reaching its highest levels
here. This phase lasts for about two weeks until
the pH decreases sufficiently to inhibit the
growth of all bacteria. The fodder mass stabili-
zes in roughly 21 days, and fermentation ceases,
assuming no air enters. If fermentation is suc-
cessful during this stage, the anticipated pH for
silage will range from 4.5 to 3.5, depending on
the moisture level of the forage [12; 60]. When
the conditions in the silo are not ideal, it creates
an environment conducive to the growth and ac-
tivity of clostridia, which are harmful bacteria.
These anaerobic bacteria generate butyric acid.
Both clostridia and coliforms have the ability
to decompose amino acids, transforming them
into volatile fatty acids, amines, and ammonia
[35; 74].
Silage fermentation occurs naturally in ana-
erobic environments thanks to the bacteria
found in plants, but the efficiency of this fer-
mentation can vary based on the specific types
of lactic acid bacteria present in the feed. Fac-
tors such as the speed of pH decline, the quanti-
ty of sugar that remains unutilized by bacteria,
the preservation of true protein, and the levels of
lactic acid, acetic acid, and ethanol all influence
the quality of the silage [31; 50].
Effective factors in silage preparation
The effective preservation of fodder and
other products relies on suppressing microbial
activity, particularly that of bacteria. Ideal con-
ditions for storing silage involve eliminating air
from the silo and ensuring anaerobic conditions.
Under these circumstances, the population of
lactic acid bacteria increases as they utilize the
plant’s internal sugars, producing significant
amounts of acid that lower the pH to around 4.
Preserving fodder can be challenging because it
can create favorable conditions for the growth
of harmful bacteria like clostridia. When har-
vested, the pH of the forage ranges from 6 to 7,
but with proper fermentation during ensiling,
the pH can drop to 4 or lower [35; 74].
The pH of silage drops as lactic acid and
other organic acids are generated by lactic acid
bacteria. Accelerating the pH reduction is cru-
cial for preserving the nutritional quality of the
silage. A decrease in pH also leads to a reduc-
tion in the populations of clostridia and coli-
form bacteria [35; 82]. In addition to reducing
the pH and inhibiting microbial activity, other
factors are also effective in preserving silage,
such as the way of filling the silo, the size of the
chopped fodder, proper compression of fodder,
and proper structure. Silage and proper silage
closure are noted [12; 60; 135].
Effective microorganisms in the ensiling
process
The bacteria that are typically sought after
in the ensiling process belong to the lactic acid-
producing group, which includes species such as
Streptococcus, Lactobacillus, and Pedicococ-
cus, among others (Fig. 6) [20;103].
These bacteria generate lactic acid by fer-
menting water-soluble carbohydrates and can
utilize organic acids like citric and malic acid
found in plant cells as a substrate for mixed
fermentation. A high concentration of water-so-
luble carbohydrates enhances the activity of lac-
tic acid-producing bacteria. Conversely, a low
concentration of these carbohydrates promotes
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 40.
9
Fig. 6. Main groups of microorganisms involved in the fermentation process [135] .
the activity of clostridial bacteria, which are un-
desirable as they thrive in anaerobic environ-
ments and lead to silage spoilage by producing
butyric acid. These unwanted bacteria compete
with lactic acid producers for water-soluble car-
bohydrates and yield less acidic byproducts
such as acetate, butyrate, propionate, and etha-
nol. Clostridia growth is more pronounced in
plants with high moisture content but is sup-
pressed at a pH of 4 or lower, which also inhi-
bits most plant protease activity [12]. The main
objective of using silage additives is to promote
the growth of lactic acid bacteria during fermen-
tation, leading to the creation of high-quality si-
lage. These additives facilitate the desired fer-
mentation process, restrict unwanted fermenta-
tion, and enhance the nutritional value of the si-
lage [20; 103].
Silage additives can be classified into vari-
ous groups: Fermentation stimulators, which in-
clude microbial inoculants like enzymes (cellu-
lase, hemicellulase) and molasses; fermentation
inhibitors, such as formic acid, formaldehyde,
and sulfuric acid; nutrients like ammonia and
urea; moisture-absorbing substances; and agents
that prevent aerobic spoilage [139]. The micro-
bial mass that can be used consists of both dry
and living microbes, which remain inactive until
they are hydrated. This microbial material is
available in two forms: liquid and dry. For the
liquid form, the microbes need to be activated
before they interact with the fodder to minimize
their dormant period and maximize their fer-
mentation capability. In the dry form, the mic-
roorganisms are activated by the moisture in the
fodder, resulting in a slower initial fermentation
rate. It is important to follow the manufacturer’s
guidelines when using these products [23].
The most commonly used microbes for mi-
crobial impregnation are lactic acid bacteria,
such as Lactobacillus, Epenococcus, and Ente-
rococcus species. These bacteria enhance lactic
acid production, leading to a quicker drop in pH
and inhibiting the growth of undesirable fer-
mentation microorganisms in silage. The high
natural bacterial populations in silage can hinder
microbial impregnation from achieving a com-
petitive edge. In contrast, alfalfa and clover
have lower natural bacterial levels, making their
microbial inoculation more effective than that of
corn silage [23]. Kung et al. [72] discovered that
microbial impregnation improved the fermenta-
tion process in both grass and legume silage.
The process of microbial impregnation in
silage preparation aims to lower the pH in the
early phases of fermentation, maintain plant
carbohydrates through uniform fermentation,
and protect plant protein by minimizing prote-
olysis and deamination [36; 50; 116]. As a re-
sult, we anticipate that treated silage will en-
hance animal performance [83]. The second
type of additives consists of fermentation inhibi-
tors that prevent undesirable fermentation.
These products typically contain a mix of acids.
The goal of using these acids is not to halt the
fermentation process, but rather to enhance the
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 40.
10
natural fermentation by introducing moderate
levels of acids [116].
The importance of the alfalfa plant in
animal nutrition
Alfalfa and various other perennial legumi-
nous plants are significant forage crops due to
their capacity to generate a substantial quantity
of high-quality fodder. No other group of forage
products can offer a superior balance of energy,
protein, and minerals for high-yielding livestock
[33; 122].
Traditionally, legumes have been utilized as
natural grazing and dry feed, but in recent years,
the practice of ensiling these products has be-
come a popular method of preservation [18],
particularly in areas with high rainfall that re-
stricts the production of dry fodder. Nonethe-
less, it is important to take into account certain
adjustments to the livestock feeding strategy, as
the chemical and physical characteristics of fo-
rage can alter during the ensiling process. By ful-
ly grasping this process, it is possible to convert
nearly all agricultural products into silage with-
out negatively impacting livestock production
[25; 45]. Alfalfa can typically be fermented into
stable silage, as long as its limitations are clear-
ly recognized. Key issues with using alfalfa as
silage fodder include insufficient water-soluble
carbohydrates, a high buffering capacity, hollow
stems, and low dry matter content [52].
Final products of fermentation
The ultimate fermentation products are in-
dicative of the predominant microorganisms
present during the silage process and signifi-
cantly influence the overall quality of the silage.
Microorganisms involved in silage fermentation
facilitate the conversion of water-soluble carbo-
hydrates into organic acids. The end products of
this fermentation process can arise from various
soluble sugars [103]. Lactate serves as a marker
for bacterial fermentation characterized by the
production of uniform lactic acid during the
fermentation process. In contrast, the presence
of a combination of acetate, propionate, and lac-
tate suggests the prevalence of heterogeneous
fermentation. Elevated levels of butyrate and am-
monia in silage are indicative of fermentation by
Clostridium spp. Additionally, ammonia nitro-
gen is generated through the action of plant en-
zymes and enterobacteria, as well as through the
processes of nitrite and nitrate regeneration [52].
The effect of maturity on soluble carbo-
hydrate content
Generally, the concentration of sugars in al-
falfa diminishes as the plant matures. Raguse
and Smith [115] documented a 15.3 % reduction
in the total quantity of non-structural sugars.
Additionally, it has been observed that there is a
decline of 51.8 % in sucrose levels, 19.2 % in
glucose levels, and 14.8 % in fructose levels
from the budding stage to the 50 % flowering
stage, a phenomenon that some researchers at-
tribute to a reduction in the leaf-to-stem ratio.
Other scholars have suggested that this decline
can be ascribed to two primary factors: first, a
decrease in photosynthetic activity of the leaves
as the plant ages, and second, an increase in
shading resulting from higher plant density,
which further diminishes photosynthesis. Fur-
thermore, the stage of maturity also influences
the content of soluble carbohydrates post-har-
vest, particularly in relation to plant respiration
(Fig. 7). Geronimo and Beevers [46] indicated
that 40 % to 60 % of the reduction in respiratory
rate is mitigated by the maturation process.
Forage quality typically indicates the over-
all nutrient content that livestock can obtain
from forages. Several factors influence forage
quality, which can be divided into three catego-
ries: plant factors, animal factors, and environ-
mental factors. Gaining a clear understanding of
how these factors interact with forage quality
can assist in selecting the right forages and sup-
plements to meet animal needs, ultimately lead-
ing to improved livestock performance and eco-
nomic benefits [63]. Amer et al. [6] determined
that the nutritional value of forage sorghum si-
lage is better than that of forage millet silage
when both are harvested at the same point in
their physiological growth.
The effect of ensiling on soluble carbo-
hydrates
During the ensiling process, soluble carbo-
hydrates undergo conversion into organic acids.
It is widely recognized that lactic acid bacteria
lack the capability to hydrolyze starch during
this phase of ensiling. According to Smith
[115], a minimum concentration of 6 % to 7 %
water-soluble carbohydrates, relative to dry
matter, is necessary to achieve a silage pH
of 4. However, in practical applications, this op-
timal level of soluble carbohydrates may not be
readily available in the field. The concentration
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Fig. 7. Maturity influencing forage nutritional quality [63] .
of soluble carbohydrates in alfalfa fodder typi-
cally ranges from 2 to 7 percent of dry matter,
with variations influenced by the stage of ma-
turity and prevailing weather conditions.
Buffering capacity in silage fodder
The buffering capacity of plant materials is
closely associated with their ability to resist
changes in pH, which is a critical consideration
in the ensiling process. It is widely believed that
organic acids and their corresponding salts play
a predominant role in the buffering capacity ob-
served under silo conditions. Conversely, some
researchers propose that inorganic ions contri-
bute significantly to the buffering activity of le-
gumes. Recent studies lend support to the hy-
pothesis that organic acids are the primary con-
tributors to this buffering capacity. The concen-
tration of organic acids in grass forage typically
ranges from 2 % to 6 %, while in legume fo-
rage, it is generally higher, approaching 6 %
to 8 %. Additionally, proteins appear to have
a minimal impact on the buffering capacity of
forage materials [25; 33].
The effect of wilting on buffering capa-
city
Wilting diminishes the buffering capacity
of fodder in comparison to fodder that is ensiled
immediately, a phenomenon attributed to the
transformation of organic acids into water and
carbon dioxide during the respiration process.
The rate of water loss during the wilting stage
influences the quantity of residual organic acids,
subsequently affecting the buffering capacity;
a gradual loss of water can further deplete the
levels of remaining organic acids. Research in-
dicates that the buffering capacity declines by
approximately 150 % as the maturity stage pro-
gresses from pre-budding to 50 % flowering
[91; 126].
Effect of wilting on silage quality
Wilting is applied to optimize the amount
of dry matter of forage before ensiling. Silage
with very low dry matter is often associated
with increased effluent production and clostridi-
um fermentation. Whereas, silage with high dry
matter does not compress well and greatly re-
duces aerobic stability [33]. The optimal dry
matter for ensiling alfalfa depends on the type
of silo construction, environmental and mana-
gement conditions. Ishler et al. [57] suggested
that the amount of dry matter suitable for silage
is 30 to 35, 40 to 45, 45 to 60 percent for tower,
oxygen-free and bag silos, respectively [113].
The moisture content of ensiled plants sig-
nificantly influences both the total bacterial
population and the rate of fermentation. The
process of wilting tends to inhibit bacterial pro-
liferation, whereas the addition of water to fo-
rage promotes bacterial growth, particularly
among lactobacilli and gram-negative bacteria.
Furthermore, wilting may impact the relative
proliferation of both homogeneous (homofer-
mentative) and heterogeneous (heterofermenta-
tive) lactic acid bacteria. Generally, silages of
inferior quality are produced under two specific
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conditions: first, when silages possess low dry
matter content, which diminishes ammonia pro-
duction due to clostridial fermentation; and se-
cond, when silages exhibit excessively high dry
matter content. In the latter scenario, although
fermentation activity is reduced, there is a pro-
nounced increase in the growth of yeasts and
fungi, leading to elevated heat production. This
heat generation subsequently contributes to an
increase in the formation of insoluble nitrogen
in acid detergent (ADIN) and results in the deg-
radation of certain amino acids, including me-
thionine, cysteine, and tyrosine [127]. McDo-
nald et al. [83] conducted a study examining the
qualitative alterations in lactobacilli during the
ensiling process of gramine and red clover fo-
rage under varying humidity conditions. Their
findings indicated that after a 142-day ensiling
period, 75 % of the total lactobacilli present in
silage with high humidity and 98 % of those in
silage with low humidity were classified as he-
terogeneous types.
Wilting elevates the energy demands asso-
ciated with the maintenance and growth of mi-
crobial populations, while also prolonging the
digestion of plant cells. Additionally, it dimini-
shes plant respiration and enzymatic activity.
Notably, although wilting does not decrease the
overall level of proteolysis, it has the potential
to mitigate this process under optimal condi-
tions [4; 22]. Owens et al. [105] found that ferti-
lization not only failed to decrease the levels of
non-protein nitrogen but actually resulted in an
increase. The process of wilting the plant inhi-
bits the release of plant effluent, which is signi-
ficant for two reasons: first, it avoids the pollu-
ting effluent; second, the leachate released in
this manner affects both the environment and
the nutritional quality of silage [48].
The research conducted by Whiter and
Kung [132] demonstrated that lactic acid-pro-
ducing bacteria exhibit greater resistance to va-
riations in osmolarity compared to other micro-
bial species and are relatively unaffected by
humidity levels. Their findings revealed that the
ratio of lactic acid to acetic acid in dry matter at
30 % concentration was between 2.6 % and
3 %, while at 54 % dry matter concentration,
this ratio exceeded 8.5 %. This observation sug-
gests a notable increase in the ratio of homoge-
neous to heterogeneous fermentation processes.
Kung et al. [71] conducted a study on alfal-
fa fodder with varying dry matter content of
30 %, 40 %, 50 %, and 60 %. Their findings in-
dicated that the pH level increased at 60 % dry
matter concentration. Additionally, they obser-
ved a reduction in lactic acid levels when dry
matter exceeded 40 %, while the concentration
of remaining soluble sugars increased. This phe-
nomenon was attributed to the stimulation of
microbial growth due to higher moisture levels,
which in turn led to an increased consumption
of soluble carbohydrates. Furthermore, to estab-
lish stable conditions during the silo’s stable
phase, a greater production of lactic acid was
necessary to inhibit bacterial activity. The study
also revealed a general decline in crude protein
content with increasing dry matter, which was
linked to leaf loss during the harvesting and si-
lage processes. Moreover, the wilted treatments
exhibited a decrease in dissolved nitrogen, am-
monia nitrogen, and free amino acids, which
was associated with reduced clostridial and en-
zymatic activity. Gou et al. [49] studied the op-
timal harvest timing for ensiling alfalfa by col-
lecting first and second cuts at the budding stage
(BS), initial flowering stage (IFS), and full
flowering stage (FFS) during 2016 and 2017.
After harvesting, the alfalfa was ensiled for
45 days. Their findings indicated that, in terms
of nutritional quality, fermentation properties,
and in vitro rumen digestibility, the ideal har-
vest stages were BS for the first cut and IFS for
the second cut. Additionally, they found that al-
falfa silage from the first cut was superior to
that from the second cut at the same maturity
stage.
Wilting has been shown to enhance the
concentration of residual soluble carbohydrates
in comparison to non-wilted forage through
several mechanisms. Firstly, the process of wil-
ting decreases water activity, which in turn in-
hibits microbial growth [28]. Additionally, du-
ring the wilting process, a significant number of
bacteria are eliminated relative to unharvested
fodder, thereby limiting fermentation [71]. Con-
sequently, the introduction of bacteria to wilted
forage appears to be essential. Furthermore,
wilting reduces the buffering capacity of the
forage, which diminishes the quantity of lactic
acid required to establish stable conditions, ul-
timately preventing further fermentation by
maintaining these stable conditions [33].
In the study conducted by Whiter and Kung
[132], alfalfa was evaluated at two distinct lev-
els of dry matter content, specifically 30 % and
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54 %, utilizing two varieties of soluble and
dried microbial additives. The findings indica-
ted a significant decline in the proliferation of
lactic acid bacteria within the silage characte-
rized by higher dry matter on the second day,
with a return to baseline levels observed by the
eighth day. Notably, no significant differences
were detected between the two experimental
groups after the fourteenth day. Furthermore,
the concentration of acetic acid in the dry matter
remained unaffected by the introduction of bac-
terial additives. This experiment revealed that
acetic acid-producing bacteria exhibit activity
during the initial phases of ensiling, and their
growth is not inhibited in less moist fodder,
where the pH decreases at a more gradual rate,
indicating that the additives do not influence
their levels. Additionally, it was observed that
the population of lactic acid bacteria, such as
Lactobacillus plantarum, diminishes when water
activity decreases from 0.987 to 0.949. Ridla et
al. [111] conducted a meta-analysis to evaluate
the impact of wilted and unwilted silage on va-
rious parameters. They demonstrated that wil-
ting prior to ensiling greatly enhanced silage
quality by boosting dry matter and water-solub-
le carbohydrates, while also decreasing dry mat-
ter losses, butyric acid, and ammonia levels.
Notably, wilting did not significantly affect pH,
crude protein, or in vitro dry matter digestibility.
Silage epiphytic population
These microorganisms are inherently found
on the fodder itself, with lactic acid bacteria, en-
terobacteria, Clostridium species, yeasts, fungi,
and aerobic bacteria being particularly signifi-
cant in determining the quality of silage. The
diversity of aerobic bacteria is influenced by
various factors, including plant species, diffe-
rent plant parts, climatic conditions, seasonal
variations, the process of wilting, and mechani-
cal crushing [49; 51]. This group of microorga-
nisms requires aerobic respiration, utilizing mo-
lecular oxygen as an oxidizing agent to generate
the energy necessary for their metabolic pro-
cesses. Aerobic bacteria are prevalent during the
growth phase of plants and continue to engage
in fermentation and respiration for several hours
post-harvest and ensiling. They remain metabo-
lically active and are capable of utilizing over
100 different types of organic compounds,
which can lead to a reduction in nutritional va-
lue by approximately 1 % to 2 % during the ini-
tial stages of the ensiling process (Table 1)
[134].
This issue can be mitigated by promptly es-
tablishing an anaerobic environment. Upon the
opening of the silo, microbial activity resumes,
leading to aerobic degradation [33]. Research
conducted by Woolford [134] identified bacilli
as the primary agents of aerobic decomposition
during the feeding phase. However, more recent
studies have indicated that certain species of
acetobacter also play a crucial role in initiating
this decomposition process, particularly under
low pH conditions where they metabolize fer-
mentation byproducts. This metabolic activity
not only diminishes the nutritional quality of si-
lage but also poses a potential health risk to
livestock and individuals handling these materi-
als due to the production of endotoxins [134].
Enterobacters
This group encompasses a diverse family
of bacteria characterized as gram-negative, non-
spore-forming, facultatively anaerobic, and
Table 1. The substrate being fermented by epiphytic microorganism [22; 83]
Organism Substrate Products
LAB (Ho) Glucose 2 lactate
LAB (He) 3 Fructose 1 lactate, 1 acetate, 2 mannitol, 1 CO
Enterobacteria 2 Glucose 1 lactate, 1 acetate, 1ethanol, 2 CO2
LAB (He) Glucose lactate, 1 ethanol, 1 CO2
LAB (Ho/He) 2 Citrate 1 lactate, 3 acetate, 3 CO2
LAB (Ho/He) Malate 1 lactate, 1 CO2
Yeasts Glucose 2 ethanol, 2 CO2
Clostridia 2 Lactate 1 butyrate, 2 CO2, 2 H
LAB: lactic acid bacteria; HO: homofermentative; HE: heterofermentative.
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typically motile, although some may exhibit
non-motility. These bacteria, which are rod-
shaped and metabolize soluble carbohydrates,
are commonly referred to as coliforms or acetic
acid-producing bacteria, although the latter desi-
gnation is often misapplied [33; 83]. Research
findings regarding the population dynamics of
these bacteria during placentation have been in-
consistent; some studies report an increase in
their numbers, while others indicate a decrease.
These variations are largely contingent upon the
initial population size prior to the onset of pla-
centation.
The process of chopping forage enhances
the proliferation of specific microbial popula-
tions. The abundance of these bacteria typically
equals or exceeds that of lactic acid bacteria
[25]. Under anaerobic conditions, these micro-
organisms exhibit a pronounced requirement for
fermentable carbohydrates. As lactic acid bacte-
ria proliferate and pH levels decline rapidly, en-
terobacteria experience a significant reduction
in numbers. However, if the decrease in pH is
delayed or if formic acid is introduced, these
bacteria demonstrate resilience. Enterobacteria
primarily generate acetic acid, lactic acid, car-
bon dioxide, hydrogen, and trace amounts of
ethanol and 2,3-butanediol.
The population of Enterobacteriaceae ex-
hibited an increase during the initial days of the
ensiling process, ultimately reaching a peak
concentration of 108 to 1010 CFU per gram in
both grass and leguminous fodder. Although
these bacteria possess limited proteolytic activi-
ty, they are capable of deaminating or decar-
boxylating certain amino acids, and a majority
of species are known to regenerate nitrate. En-
terobacter species are particularly notable for
their capacity to generate substantial amounts of
ammonia during ensiling, which plays a crucial
role in the decomposition of nitrate under condi-
tions conducive to regeneration within the si-
lage. This biochemical process is essential for
determining the final chemical quality of the si-
lage and serves to inhibit the proliferation of
Clostridium by facilitating the production of ni-
trate and nitric oxide derived from nitric oxide
[135; 137].
In the initial phase of the ensiling process,
there is a notable increase in the population of
enterobacteria, which coincides with a peak in
nitrate concentration. This period is characte-
rized by the maximum decomposition of nitrate,
which may serve as an effective anti-clostridial
agent of endogenous origin, particularly under
conditions of elevated pH when Clostridium
spores begin to proliferate. However, in regions
where fodder is processed into silage, the rela-
tionship between nitrate levels and the suppres-
sion of Clostridium activity remains inadequate-
ly substantiated. The significant reduction in
Clostridium activity is more commonly attribu-
ted to decreased water activity and increased
osmolality within the silage, rather than the
presence of elevated nitrate concentrations.
Generally, the presence of enterobacteria is con-
sidered undesirable, as these microorganisms
compete with lactic acid bacteria for nutrient
uptake and can produce endotoxins. Further-
more, they may contribute to ammonia for-
mation through protein degradation and nitrate
reduction during the ensiling process, which can
enhance buffering capacity and inhibit pH de-
cline. Ultimately, the rapid decline of enterobac-
terial populations serves as a more reliable indi-
cator of high-quality silage production than any
other metric [18], [135].
In the study conducted by Kizilsimsek et al.
[66], it was demonstrated that the concentration
of enterobacters in fodder was notably elevated,
corroborating the findings of Lin et al. [145].
The bacterial counts remained consistent across
treatments during the initial six hours of ensi-
ling. However, at the twelve-hour mark, a tre-
atment incorporating additives in the form of
fresh culture exhibited a reduction in bacterial
numbers, and by twenty-four hours, both types
of additives significantly decreased the entero-
bacters compared to the control treatment. Prior
research has indicated that in inoculated treat-
ments, the decline of these microorganisms oc-
curs at an accelerated pace. The authors refe-
renced Kung et al. [72] to explain these observa-
tions, attributing the results to the sensitivity of
enterobacters to low pH levels.
Clostridium
These bacteria are characterized as gram-
positive, spore-forming, typically motile, obli-
gate anaerobes, and exhibit a rod-shaped mor-
phology. They are categorized based on their
substrate utilization into two distinct groups:
1) saccharolytic bacteria, which primarily meta-
bolize sugars with minimal activity on proteins,
and 2) proteolytic bacteria, which ferment ami-
no acids. Certain species possess the capability
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to utilize both fermentation substrates [25]. In
unharvested fodder, the concentration of these
microorganisms can reach approximately 100
per gram of fresh material; however, this num-
ber significantly increases following harvesting
and processing. It is important to note that these
bacteria are not epiphytic; their presence in si-
lage is attributed to contamination from soil and
fecal matter during the harvesting process or
from precipitation entering the silage environ-
ment. Due to their obligate anaerobic nature,
these bacteria become active in the subsequent
stages of ensiling, when oxygen is absent and
anaerobic conditions prevail [33].
A pH level exceeding 5, which is conducive
to the activity of Clostridium, also provides an
optimal environment for the functioning of plant
proteolytic enzymes. The action of these en-
zymes facilitates the release of amino acids,
which Clostridium subsequently utilizes in three
distinct processes: 1) deamination, leading to
the formation of organic acids and ammonia;
2) decarboxylation, resulting in the production of
amines and carbon dioxide; and 3) ongoing re-
actions that yield organic acids, carbon dioxide,
ammonia, and alcohols [83]. Clostridium fer-
mentation enhances proteolytic activity, leading
to the generation of water-soluble nitrogen,
which can be categorized into two components:
ammonia nitrogen and non-ammonia nitrogen.
The presence of ammonia nitrogen significantly
reduces the quantity of metabolizable protein,
potentially impacting milk production, milk
urea nitrogen levels, and nitrogen use efficiency
in dairy production [52]. A direct correlation
exists between the concentration of clostridia
and the quality of silage, with significant clos-
tridial activity potentially resulting in a reduc-
tion of up to 50 % in the nutritional value of the
silage. Several factors that may influence the
proliferation of these bacteria include: 1) tem-
perature, 2) the proportion of dry matter, 3) the
level of soluble carbohydrates, 4) the buffering
capacity of the feed, and 5) the efficiency of silo
closure and compaction [70; 89].
In addition to losses incurred during fer-
mentation, spores from Clostridium species may
contaminate milk and impede the coagulation
process in hard cheese production [83]. Certain
Clostridium species, such as Clostridium butyri-
cum, are known to cause botulism in silage that
has been contaminated with soil, posing a risk to
livestock. Given that these bacteria exhibit sen-
sitivity to acidic pH levels, their proliferation
can be mitigated through the application of or-
ganic and mineral acids. However, organic acids
demonstrate greater efficacy than mineral acids
in inhibiting the growth of these microorga-
nisms. Furthermore, the incorporation of micro-
bial additives represents an additional strategy
to curtail the proliferation of these bacteria.
These additives not only facilitate the produc-
tion of lactic acid and other volatile fatty acids
but also inhibit microbial growth. Additionally,
they can produce antibiotic compounds, such as
nisin, which serves as a food preservative and
further suppresses the growth of these bacterial
species [134].
Clostridia thrive in humid environments;
however, when the dry matter content of a plant
reaches 30 % and the plant is wilted, the proli-
feration of these bacteria is inhibited. Clostridia
exhibit a higher sensitivity to water activity
compared to lactic acid bacteria [132]. Further-
more, the critical pH level conducive to Clos-
tridium growth is directly correlated with water
activity levels. Water activity, alongside dry
matter or moisture content, serves as a more
precise indicator of the water available for mi-
crobial growth during the fermentation process.
It influences all four phases of microbial
growth: 1) the duration of the incubation period,
2) the growth rate, 3) the stabilization or sta-
tionary phase, and 4) the rate of bacterial mor-
tality [121; 132].
The buffering capacity significantly influ-
ences the proliferation of these bacteria; as the
buffering capacity of the plant increases, the
quantity of lactic acid required to lower the pH
to a critical threshold that inhibits the growth of
Clostridium also rises. A primary challenge as-
sociated with legumes is their elevated buffering
capacity coupled with a low concentration of
water-soluble carbohydrates, which typically re-
sults in the dominance of Clostridium, unless
the plants are ground prior to ensiling or suita-
ble additives are employed. The relationship be-
tween temperature and Clostridium growth is
complex, as the heat generated within silos is pri-
marily due to the oxidation of sugars, leading to
a depletion of soluble carbohydrates and a sub-
sequent reduction in lactic acid production. Fur-
thermore, it appears that elevated temperatures
may promote the growth of Clostridium [102].
It is widely accepted that Clostridia are
strictly anaerobic organisms, which suggests
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they do not contribute to aerobic spoilage.
However, Jonsson [61] demonstrated that the
introduction of air can indirectly promote the
proliferation of Clostridium thyrobutyricum. In
his study, silage produced under optimal condi-
tions within laboratory silos exhibited a signifi-
cant increase in Clostridium spores and butyric
acid when exposed to air and subjected to aero-
bic degradation. This phenomenon was obser-
ved both on the surface of the silage and in
deeper regions characterized by low pH levels.
It is hypothesized that Clostridia thrive in loca-
lized microenvironments created by aerobic mi-
croorganisms, such as yeasts and certain Bacil-
lus species, which utilize lactic acid and amino
acids. This growth is likely facilitated by a re-
duction in lactic acid concentration, a decrease
in oxygen levels, and an increase in pH, ulti-
mately resulting in conditions conducive to
Clostridium proliferation.
In a study conducted by Vissers et al. [125]
to corroborate the findings of Jonsson [61], the
researchers examined the concentration of Clos-
tridium spores and its correlation with aerobic
stability. The initial experiment revealed that the
quantity of spores ingested by livestock is influ-
enced by small portions of silage that contain a
high density of spores. Elevated concentrations
of spores are typically found in regions where
mold is present. Specifically, the areas exhibi-
ting mold on the surfaces of grass silage and
corn silage account for approximately 21 % and
19 %, respectively, of the regions with elevated
spore levels. In the subsequent experiment, it
was demonstrated that the concentration of bu-
tyric acid-producing bacterial spores is predo-
minantly found within the upper 50 cm of sila-
ge, and an increase in spore numbers is associa-
ted with indicators of aerobic instability. Further-
more, high concentrations of yeasts, as well as
areas exhibiting temperatures 5 degrees Celsius
above ambient or pH levels exceeding 4.4, are
generally correlated with a significant presence
of spores. The researchers posited that the ele-
vated levels of butyric acid-producing bacterial
spores in corn silage are associated with regions
where oxygen infiltration occurs, resulting in
aerobic decomposition. This process contributes
to the formation of anaerobic conditions and an
increase in pH at the silage surface, thereby fa-
cilitating the proliferation of butyric acid-
producing bacteria in these specific areas [125].
Yeasts and molds
Yeasts are characterized as unicellular or-
ganisms, whereas molds consist of multicellular
filamentous structures. The majority of fungal
species are aerobic, requiring oxygen for their
growth; however, yeasts possess the capability
to generate energy in anaerobic environments.
The proliferation of yeasts and molds is en-
hanced by the incorporation of silage, and their
populations tend to rise following the growing
season [41]. Yeasts are known to metabolize
sugars into alcohol [33], which is linked to or-
ganoleptic quality issues in milk. This fermenta-
tion process can lead to alterations in rumen
fermentation dynamics, resulting in increased
concentrations of rumen acetate and caproic
acid, ultimately diminishing the palatability of
the feed. Furthermore, yeasts possess the capa-
bility to utilize organic acids [69] and can pro-
duce mycotoxins, rendering them an undesirable
component in silage. They play a significant
role in aerobic degradation and can be classified
into two categories: 1) lactate fermenters and
2) sugar consumers.
According to Woolford [134], the presence
of lactate-degrading yeasts at concentrations of
105 CFU per gram of material increases the like-
lihood of silage spoilage. Subsequently, molds
appear to contribute to the further degradation
process. The anaerobic and acidic conditions
characteristic of silos are highly unfavorable for
yeast proliferation. Short-chain fatty acids, such
as propionate and acetate, exhibit inhibitory ef-
fects on yeast growth, with unsaturated fatty
acids demonstrating even greater efficacy. This
enhanced effectiveness is likely attributed to the
more rapid penetration of these compounds into
yeast cells, where they subsequently lower the
intracellular pH by releasing protons, leading to
the rapid destruction of the yeasts. Under aero-
bic conditions, the energy required to extrude
protons is derived from the oxidation of various
substrates, whereas in anaerobic conditions, this
energy is obtained through the fermentation of
sugars, a process that is notably less efficient.
Most yeasts require oxygen for alcohol produc-
tion, and in its absence, the growth of many
yeast species is significantly impeded [25].
The yeasts is influenced by several key fac-
tors: 1) the volume of air that enters the silo du-
ring the ensiling process, 2) the specific type of
silage plant utilized, and 3) the application
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of silage additives. For instance, the use of for-
mic acid has been shown to enhance yeast popu-
lations, whereas bacterial additives that generate
heterogeneous lactic acid tend to diminish their
numbers [42; 131]. Additionally, research con-
ducted by Lindgren et al. [76] indicated that the
yeast populations present in alfalfa and grass si-
lages are more abundant than those of Torulop-
sis and Rhodotorula aerobic species.
Lactic acid producing bacteria (LAB)
This group possesses the capability to syn-
thesize lactic acid and was first recognized in
1900. In 1919, Oral Genus characterized true
lactic acid-producing bacteria as an epiphytic
assemblage of gram-positive, non-spore-for-
ming, non-motile, rod-shaped, and spherical
microorganisms that primarily ferment carbo-
hydrates and certain alcohols into lactic acid.
The genera of epiphytic lactic acid-producing
bacteria include Lactobacillus, Streptococcus,
Enterococcus, Pediococcus, Lactococcus, and
Leuconostoc (Fig. 8) [109].
Bacteria involved in the fermentation of
hexoses can be categorized into three distinct
groups based on their metabolic pathways. The
first group, known as compulsory homogeneous
fermenters, exclusively produces lactic acid.
The second group, referred to as optional hete-
rogeneous fermenters, primarily generates lactic
acid but also possesses the capability to ferment
pentoses into both lactic acid and acetic acid.
The third group, termed heterogeneous obligate
fermenters, is characterized by its ability to
convert hexoses into a range of products, in-
cluding lactic acid, acetic acid, ethanol, and car-
bon dioxide [33].
The population of lactic acid bacteria pre-
sent on a living plant is relatively low and is in-
fluenced by various factors, including plant spe-
cies, growth stage, environmental conditions,
seasonal variations, and the processes of plan-
ting or crushing. Notably, environmental condi-
tions, seasonal changes, and the act of crushing
appear to be the most significant determinants
of this bacterial population [109].
Muck [90] indicated that the population of
lactic acid bacteria can be estimated based on
various meteorological factors, including air
temperature, sunlight exposure, precipitation,
and relative humidity. The bacterial count on
fresh plant material ranges from 103 to 105 and
is significantly affected by the processes of har-
vesting and crushing, a phenomenon referred to
as inoculation by crushing, which can lead to an
approximate 100-fold increase in lactic acid-
producing bacteria. Notably, lactic acid bacteria
are characterized by the absence of the catalase
enzyme, rendering them incapable of detoxi-
fying peroxides. The mechanical disruption of
plant cells during harvesting and crushing re-
leases compounds such as superoxide dismutase
Fig. 8. The presence and activity of LAB in various ecological niches: A broad application
in agriculture, environmental science, and health functions [109] .
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and manganese, which become accessible to the
bacteria and exhibit effects analogous to those
of catalase.
In the early phases of the ensiling process,
the growth of lactococci and lactobacilli, in-
cluding Lactobacillus plantarum, occurs along-
side aerobic microorganisms such as yeasts,
fungi, and aerobic bacteria, facilitated by the
presence of air among the plant materials. As
fermentation advances, an anaerobic environ-
ment is established, leading to the predominance
of lactic acid-producing bacteria, particularly
Lactobacillus plantarum, which exhibit a high
tolerance to acidic conditions. It appears that the
various species identified on the plant are fre-
quently heterogeneous lactic acid-producing
species [92]. But some researchers have identi-
fied homogeneous fermenters as predominant,
highlighting the interdependent influence of
plant species, growing season, and climatic con-
ditions on the microbial composition of silage
forage plants. Recent findings further elucidate
this phenomenon, indicating that when hexose
availability is restricted, specific species of lac-
tic acid bacteria can utilize lactic acid as an ene-
rgy source to produce acetic acid under anae-
robic conditions. This metabolic process results
in an elevation of pH, which subsequently fos-
ters the proliferation of microorganisms, inclu-
ding non-beneficial organisms such as Clostri-
dium and Enterobacter [92]. Lactic acid bacteria
exhibit proteolytic capabilities; however, their
action on amino acids appears to be restricted.
Furthermore, it is believed that certain lactic
acid bacteria are capable of fermenting two spe-
cific amino acids, namely serine and arginine.
Additionally, some bacterial species, including
Lactobacillus plantarum, possess the ability to
reduce nitrate to ammonia or nitrogen oxide
[128].
Propionic acid producing bacteria
These bacteria serve as a natural source for
the production of propionic acid. Their advanta-
geous properties, including the synthesis of pro-
pionic acid, bacteriocins, and vitamin B12, as
well as their capacity to thrive and proliferate in
the rumen, have led to their application as fer-
mentation enhancers in silos and in high-
moisture food products. Furthermore, these bac-
teria possess a diverse array of peptidases, ena-
bling them to degrade various amino acids;
however, significant variations exist among dif-
ferent strains [76].
Propionic acid is extensively utilized in the
preservation of seeds, dry matter, fodder, and
silage. Research conducted by Moon et al. [88]
demonstrated the synergistic effects of acetic
acid, lactic acid, and propionic acid in comba-
ting acid-resistant yeasts. In addition to propio-
nic acid, certain bacteria are known to produce
antimicrobial proteins, referred to as bacterio-
cins. These bacteria exhibit stability at tempera-
tures below 85 degrees Celsius and within a pH
range of 3 to 9. The presence of propionic acid
and other volatile fatty acids inhibits cellular
growth, which may be attributed to their sup-
pressive effects on the absorption of amino
acids and other essential compounds necessary
for cellular proliferation, as well as their role in
obstructing ATP production, which relies on the
electron transport chain.
Contrast between lactic acid and propi-
onic acid producing bacteria
Lactobacilli are capable of synthesizing lac-
tic acid from sugars, while propionic acid-
producing bacteria generate propionic acid from
both sugars and lactic acid. The extent to which
lactic acid producers stimulate propionic acid
producers is contingent upon the specific bacte-
rial strain involved. Research conducted by Pe-
rez-chaia et al. [106] identified an inhibitory ef-
fect of lactic acid-producing bacteria on propi-
onic acid production, which was attributed to a
rapid decline in pH levels. Furthermore, lactic
acid-producing bacteria generate both D and
L isomers, whereas propionic acid-producing
bacteria exhibit a preference for L-lactate over
D-lactate, which serves as a stimulant for their
activity. The interaction between propionic acid-
producing and lactic acid-producing bacteria
extends beyond the dynamics of lactate produc-
tion and consumption. Lactobacilli in mixed
cultures derive advantages from the metabolic
activities of propionic bacteria, particularly due
to the production of carbon dioxide. Additional-
ly, the proteolytic activity of lactic acid bacteria
leads to the release of amino acids, which en-
hances the growth conditions for propionic acid-
producing bacteria. However, it is important to
note that excessive proteolysis resulting in an
overabundance of amino acids can inhibit the
growth of propionic acid-producing bacteria.
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Observations indicate that these mutual interac-
tions between the two bacterial species predo-
minantly occur under conditions of low glucose
and soluble sugar availability in the environ-
ment [56; 128].
In this context, lactic acid-producing bacte-
ria exhibit an inhibitory effect on other lactic
acid-producing bacteria. This phenomenon is
likely attributable to the infiltration of propionic
acid into the cells of lactic acid-producing bac-
teria, resulting in the release of protons within
the cellular environment. In an effort to mitigate
this issue, the cells attempt to extrude protons
via the H+ATPase pump, a process that necessi-
tates energy expenditure. However, the limited
availability of soluble sugars to supply the re-
quisite energy leads to a decline in both the
growth rate and biomass production of lactic
acid-producing bacteria [106]. It is important to
note that the growth constraints faced by these
bacteria, compounded by their low tolerance to
acidic conditions, have contributed to a reduc-
tion in their application as silage additives [110;
129].
Effect of microbial additive on silage
forage
The application of bacterial additives to en-
hance fermentation processes in silage has
reached a historical peak (Table 3). Currently,
a diverse array of these products, featuring vari-
ous bacterial formulations, is available in the
commercial market for this specific purpose.
The quality of silage fermentation is contingent
upon the size, diversity, and activity of the lactic
acid-producing bacterial population present in
the fodder. This population typically ranges
from 10 to 102 colony-forming units per gram of
material, escalating to levels of 106 or 107 du-
ring the harvesting and crushing phases. Never-
theless, a significant proportion of these bacteria
are heterogeneous and predominantly belong to
the Leuconostoc species, which may not be the
most effective organisms for facilitating do-
minant fermentation processes (Fig. 9, 10) [50;
95].
Since 1950, homogeneous lactic acid bacte-
ria have been effectively utilized as additives in
silage. These bacteria are added to support the
growth of the epiphytic bacterial population that
generates lactic acid, allowing them to outcom-
pete other microbes. This leads to an increased
production of lactic acid in a shorter timeframe,
which shortens the fermentation process by
minimizing proteolysis and the formation of
volatile organic acids and ethanol, ultimately
enhancing feed preservation [66].
Over the past decade, advancements in the
selection and upkeep of bacterial strains have
resulted in the creation of commercial products
that contain a sufficient quantity of uniform lac-
tic acid-producing bacteria, with a minimum
concentration of 106. This level is essential in
fresh grass to promote the prevalence of lactic
Fig. 9. Role of lactic acid bacteria on silage production and preservation [116].
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Fig. 10. Comparison structure of silage microbiome at genera level between control and
inoculant addition [112].
Table 3. Effect of LAB inoculants on silage quality
Silage type LAB inoculant Additive
composition Effect on silage preservation Refe-
rences
Alfalfa L. buchneri orange pulp
LAB can improve the aerobic stability
and quality of silage in laboratory silos
and also, OP and LAB might improve
silage quality and cause better silage
management in the farm.
[15]
Alfalfa L. buchneri orange pulp
Supplementation silage with orange
pulp and bacterial inoculant increased in
vitro DM digestibility in all incubation
times. There is highly relationship be-
tween in situ and in vitro DM digesti-
bility.
[14]
Alfalfa L. plantarum
(1 × 106 CFU/g)
cellulase
(20 mg kg–1)
L. plantarum showed the highest in
vitro digestibility of dry matter and
highest abundance of natural LAB
compared to cellulase-treated silage.
Overall silage quality of alfalfa was im-
proved.
[75]
Alfalfa
L. buchneri,
L. plantarum,
L. buchneri +
L. plantarum
(1 × 106, 2 × 106 CFU/g)
–
Decreased pH, increased the production
of lactic and acetic acids, reduced the
number of yeasts and molds, inhibited
Enterobacterium and Klebsiella pneu-
moniae, stabilized silages during aero-
bic exposure.
[143]
Lucerne, oat,
sorghum,
Whole-crop
corn
L. plantarum,
Limosilacto-bacillus
fermentum
(1 × 106 CFU/g)
–
Silages were well preserved based on
their pH and DM values, enhanced
aerobic stability of maize silage. [107]
Alfalfa L. buchneri
(3×108 CFU/g)
molasses,
orange pulp
Supplementation treatments with inocu-
lant had a significant effect on gas pro-
duction and increased gas production
volume
[17]
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Continuation of table 3
Silage type LAB inoculant Additive
composition Effect on silage preservation Refe-
rences
Alfalfa
B. subtilis,
L. buchneri
(1 × 106 CFU/g)
–
Inhibited the growth of Enterococcus
after 3 d of aerobic exposure, improved
silage fermentation quality, aerobic sta-
bility, and bacterial community during
ensiling.
[10]
Alfalfa
L. plantarum,
L. casei,
E. faecium,
P. acidilactici
(1 × 105 CFU/g)
–
Increased the organic acids content,
bacterial species number, and relative
abundances following fermentation, re-
sulting in a general pH and mycotoxin
reduction.
[11]
Alfalfa,
Chinese
rye grass
B. amyloliquefaciens,
B. licheniformis,
B. subtilis,
Paenibacillus
xylanexedens,
B. cereus, B. flexus,
E. faecium, B. pumilus
–
With a low pH and high lactic acid con-
centrations, the silages were successful-
ly preserved, loss of water-soluble car-
bohydrates, starch, and hemicellulose in
both silages with prolonged ensiling,
activity of microbial amylase was de-
tected all through the ensiling process.
[99]
Alfalfa L. buchneri fresh whey
(3 %)
Fresh whey and bacterial inoculation
with the fermentable carbohydrates
leads to rapid reduction of pH, limiting
the proliferation of yeasts, increased
aerobic stability and improved silage
quality.
[16]
Alfalfa L. plantarum
(1 × 105 CFU/g) –
Better silage quality-lower pH, greater
lactate-to-acetate ratio, higher ruminal
levels of L. plantarum and, in some
cows, a detectable shift in bacterial
community composition
[86]
Alfalfa
L. plantarum,
L. buchneri
(1 × 106 CFU/g)
–
Hastened the inhibition of E. coli during
ensiling, prevented the growth on silage
contaminated with the pathogen after
ensiling.
[100]
Alfalfa L. plantarum
(1 × 105 CFU/g) – Higher lactic acid and acetic acid con-
centrations. [29]
Alfalfa
L. brevis, L. citerum,
L. bifermentans,
L. plantarum
(1 × 105 CFU/g)
–
Positively affected fermentation proper-
ties and silage quality parameters. [40]
Alfalfa L. plantarum
(1 × 105 CFU/g) – Improved ruminal fermentation and
milk production. [87]
Alfalfa,
corn straw
P. pentosaceus,
P. acidilactici,
L. acidophilus,
L. plantarum
(2 × 105 CFU/g)
–
Improved silage characteristics and fi-
ber degradation in alfalfa, while having
no detectable effect on corn silage. [30]
Alfalfa
L. plantarum,
L. pentosus,
P. pentosaceus
(1 × 105 CFU/g)
–
Efficiently regulated enterobacteria and
mold populations, the chemical proper-
ties of the silage were improved by an
increased index of in vitro dry matter
digestibility.
[97]
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End of table 3
Alfalfa
L. pentosus,
L. pentosus +
L. brevis +
P. acidilactici
(1 × 106 CFU/g)
–
Presented greater residual WSC and the
least pH.
[140]
Corn hybrid
L. buchneri
(4 × 105 cfu/g),
P. pentosaceus
(1 × 105 CFU/g)
molasses
(3 %)
Molasses increased ethanol and lactate
concentration but did not improve aero-
bic stability, while LAB inoculants
made the fermentation more heterolactic
and improved corn silage’s aerobic sta-
bility
[55]
Rice straw L. plantarum
(1 × 106 CFU/g)
molasses
(4 %)
Molasses improved rice straw silage’s
fermentation quality and in vitro diges-
tibility.
[144]
Wilted rice
straw
L. bulgaricus +
L. helveticus
(1 × 106 CFU/g)
Acetic acid
(5 %) +
molasses
(40 %)
Treatment with chemical additives in-
creased the concentrations of CP, WSC,
acetic acid, and lactic acid reduced the
concentrations of ADF and NDF but did
not effectively inhibit the growth of
spoilage organisms as seen in the treat-
ment with LAB
[98]
Whole-crop
maize
L. plantarum,
L. paracasei,
P. pentosaceus
(1.5 × 1011 CFU/g)
Formic acid
(42.5 %),
propionic acid
(10.0 %),
ammonium
formate
(30.3 %),
benzoic acid
(2.2 %)
The concentrations of WSC were higher
for chemical additives compared to
LAB on days 3, 5, 10, and 90 of fer-
mentation, and silage had fewer LAB
populations than LAB treated silages
regardless of the days of fermentation.
[123]
Whole-crop
corn
L. acidophilus +
L. plantarum
(1 × 106 CFU/g)
Formic acid,
acetic acid,
propionic acid
(6 mLg–1)
Silages treated with LAB showed in-
creased lactic acid content and de-
creased pH after 45 days, while higher
levels of acetic acid and increased
abundance of Acetobacter in silages
treated with organic acids
[58]
acid bacteria in silage, ultimately enhancing
animal performance. The majority of microbial
additives currently on the market consist of
selected strains of uniform lactic acid-producing
bacteria, including species such as Lactobacil-
lus plantarum, Pediococcus, and Enterococcus
[25].
In general, inoculants are chosen for their
effectiveness in quickly reducing the pH of si-
lage by fermenting water-soluble carbohydrates
into lactic acids, which helps to inhibit proteo-
lytic activity and preserve nutrients. Currently,
efforts are being made to develop functional ino-
culants that not only enhance silage quality but
also have beneficial effects on animal health,
production, stress resilience, and improve silage
consumption and digestibility. Some research
has shown that silages treated with inoculants
can lead to higher milk production in cows
compared to those without inoculants [79; 84],
although the exact mechanisms behind this are
still not fully understood. In this section, we will
discuss advancements related to several promi-
sing functional inoculants for silage. These ino-
culants enhance silage safety while preserving
fermentation quality and lowering pH. Addi-
tionally, they boost animal performance, in-
crease feed intake and digestibility, and can
even improve the quality of animal products
(Fig. 11).
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23
Fig. 11. Functions of lactic acid bacteria and their effects on silage and ruminants [50] .
Factors affecting the activity of microbial
additives
Temperature
Elevated temperatures are the primary ca-
use of poor-quality silage, as the heat generated
during fermentation is positively linked to mi-
crobial growth and plant respiration. Initially,
there is a rapid increase in temperature, which
halts the growth of various bacteria. Under these
circumstances, commercial additives that con-
tain lactic acid-producing bacteria typically of-
fer minimal advantages [142]. The water tem-
perature in the inoculation tank for silage plays
a crucial role in the survival of bacterial addi-
tives, with high temperatures potentially leading
to heat shock in the bacteria [7]. In a study by
Mulrooney et al. [93], five bacterial additives
were cultured and examined at four different
temperatures over a period of six hours. The
findings indicated that the viability of the bacte-
rial additives remained stable at temperatures
between 30 and 40 °C, but temperatures excee-
ding 40 °C resulted in a decline in their viability.
Different additives exhibited varying responses
to temperature; Lactobacillus plantarum de-
monstrated the highest tolerance to elevated
temperatures, while Lactobacillus buccaneri
showed moderate tolerance. The other bacterial
strains were unable to withstand the higher tem-
peratures, leading to a significant reduction in
their numbers.
Teixeria et al. [120] found that Lactobacil-
lus buccaneri is compromised at 62 degrees due
to damage to its cell wall, and at temperatures
exceeding 65 degrees, ribosomes are affected.
Additionally, Spinks [117] noted that certain
bacteria exhibit heightened heat sensitivity du-
ring starvation when there is a scarcity of fer-
mentable substrates. The findings from these
studies indicate that heat sensitivity in lactic
acid bacteria can significantly impact the effec-
tiveness of additives in silage, as it not only en-
hances microbial survival prior to ensiling but
also improves their survival rate during the en-
siling process when temperatures rise. Ohmomo
et al. [101] noted that the poor quality of silage
and the ineffectiveness of commercial additives
may be due to temperatures exceeding 45 de-
grees during the early stages of ensiling in the
silage stack. Zhang et al. [142] found that
among eight strains of epiphytic bacteria tested
at 25 degrees, all enhanced fermentation; how-
ever, at 45 degrees, only one additive was effec-
tive in improving fermentation. Ultimately, it is
suggested that to achieve better outcomes, heat-
resistant species should be used, along with ice
packs and pure, chlorine-free water during ino-
culation.
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Preparation before use
The use of dry and liquid additives can also
affect the type of fermentation. In Whiter and
Kung’s [132] experiment, Lactobacillus planta-
rum was used in two forms: dry granules and
soluble in water on two levels of dry matter. In
low dry matter, both additives had equal per-
formance, but with increasing dry matter, silage
containing soluble additive showed its effect
better. In the study conducted by Merry et al.
[85], the use of a fresh bacterial culture com-
bined with fast-fermenting foods in a liquid en-
vironment enhanced the fermentation process
more effectively than when a freeze-dried bacte-
rial additive was applied for silage inoculation.
This improvement is likely due to a reduction in
the lag phase and a quicker increase in bacterial
population.
In the study by Kizilsimsek et al. [66], two
forms of a bacterial additive — one dried
through freezing in both high and low doses,
and another as a fresh culture — were applied to
alfalfa fodder. Within the first 6 and 12 hours
after ensiling, the number of lactic acid bacteria
and the level of lactic acid rose in the treatment
with the fresh culture. After 12 hours, levels of
acetic acid and ethanol decreased, and by 24
hours post-ensiling, ammonia nitrogen levels al-
so dropped compared to other treatments. The
findings indicated that the fresh culture bacterial
additive resulted in a quicker pH reduction and
more effective fermentation than the dried addi-
tive, which was rehydrated in water. The silage
with the fresh culture additive exhibited the
most favorable response and a more uniform
fermentation process. In their 1998 experiment,
Winters et al. [133] demonstrated that introdu-
cing fresh cultures of bacteria led to greater dry
matter consumption and weight gain compared
to the control group. Additionally, the study
highlighted that the quantity of dry matter influ-
ences the effectiveness of the bacterial additive,
particularly in relation to the pasteurization of
feed.
The effect of bacterial additives on fer-
mentation characteristics of silage
Up until now, various bacterial additives
that generate uniform lactic acid have been
applied to alfalfa silage, and their beneficial
impacts on fermentation quality have been stu-
died. In a study by Cai et al. [26], two distinct
strains of Lactobacillus plantarum were utilized
at a concentration of 105 for three alfalfa plants
with 45 % dry matter, along with rye fodder at
the flowering stage and sorghum fodder at the
milking stage. The use of the bacterial additive
notably enhanced the fermentation parameters,
leading to increased production of lactic acid
and soluble carbohydrates, as well as higher
levels of butyric acid, propionic acid, ammonia
nitrogen, and dry matter loss. Muck et al. [89]
inoculated alfalfa forage with a blend of lactic
acid bacteria and found that there was a gradual
enhancement in the rate of silage acidification
and an increase in the ratio of lactate to acetate.
They concluded that lactic acid bacteria should
be applied at levels of 10 % or higher than
the natural amounts of epiphytic lactic acid bac-
teria.
Hristov and McAllister [54] inoculated en-
tire barley plants with a combination of Entero-
coccus facium and Lactobacillus plantarum.
They found that the quantity of lactic acid bac-
teria and the total lactic acid produced rose,
while the final pH of the silage dropped in the
inoculated group. However, they concluded that
the bacterial additive was more effective when
the dry matter content was low. Koc et al. [68]
investigated the effects of two types of bacterial
additives and enzymes, finding that the inclu-
sion of bacteria enhanced fermentation parame-
ters. In treatments where both enzymes and bac-
terial additives were used simultaneously, there
was a notable decrease in pH. This reduction in
pH was accompanied by a decrease in soluble
sugar levels. Treatments with additives showed
an increase in lactic acid bacteria and lactic acid
production, while acetic acid levels were lower,
and butyric acid was not detected in any of the
treatments.
In the experiment conducted by Kung et al.
[71], the use of a bacterial additive in both nor-
mal and double doses in feed with 42 % dry
matter resulted in a reduction of pH, with no
significant difference observed between the two
doses. Initially, the soluble carbohydrates in wa-
ter were consistent across all treatments, but by
the end of the experiment, the control treatment
had a higher amount of these carbohydrates,
while lactic acid levels exhibited a negative cor-
relation with soluble carbohydrates. The fin-
dings indicated that at elevated dry matter le-
vels, pH decreased and lactic acid increased.
Ultimately, it was concluded that adding bacte-
ria to alfalfa silage led to an increase in lactic
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25
acid, with this effect being more pronounced at
higher dry matter levels.
In a separate study by Filya et al. [43], re-
searchers examined the impact of the bacterial
additive Propionibacterium, both with and with-
out Lactobacillus plantarum, on corn plants.
The treatments that included Lactobacillus
plantarum resulted in higher levels of lactic
acid, while Propionibacterium did not lead to
an increase in propionic acid. Throughout the
entire experiment, the levels of soluble carbo-
hydrates, propionic acid, acetic acid, butyric
acid, and ethanol remained unchanged due to
the bacterial additive. In the experiment con-
ducted by Rizk [113], alfalfa with a high dry
matter content was examined with the inclusion
of uniform lactic acid bacteria (Lactobacillus
plantarum). The pH levels in the treatments
with bacterial additives dropped quickly, falling
below 4.5 within two days, whereas the control
treatment only reached this pH after 45 days. By
the end of the study, the pH and soluble carbo-
hydrates in the treatment with the bacterial addi-
tive were lower than those in the control treat-
ment, but the production of lactic acid was
higher in the bacterial additive treatment.
In the study conducted by McAllister et al.
[80], two varieties of bacterial additives were
utilized, one containing Lactobacillus plantarum
and the other containing both Lactobacillus
plantarum and Enterococcus faecium. The qua-
lity of all silages was found to be good. The
presence of lactic acid bacteria showed a ten-
dency to rise, leading to an increase in lactic
acid concentration, while pH levels, butyric acid,
and water-soluble carbohydrates decreased in
the treatments with bacterial additives. The le-
vels of ammonia nitrogen remained unchanged,
but the amount of acetic acid increased by over
50 % in the treatment that included amphoteric
acid. In a study by Nadeau et al. [96], alfalfa
and orchard grass were ensiled with dry matter
contents of 22 % and 32 %, respectively. The
researchers utilized bacterial additives such as
Lactobacillus plantarum and Pediococcus cere-
visiae. The addition of a bacterial additive con-
taining cellulase to alfalfa resulted in a slight
but significant reduction in pH, a change not ob-
served in orchard grass. The presence of bacte-
ria led to an increase in lactic acid levels in both
types of fodder. The lactate to acetate ratio rose
by 15 % in alfalfa and by 8 % in orchard grass.
While the level of acetic acid in orchard grass
remained unchanged by the additives, it de-
creased by 16 % in alfalfa compared to the con-
trol group. Additionally, succinic acid levels
were higher in wilted fodder than in non-wilted
forage for both plant species, likely due to in-
creased proteolysis in the silage, which is partly
facilitated by enterobacters that convert glucose
into succinate. The study highlighted that va-
rying bacterial additive contents led to different
responses, ultimately influencing the quality of
the silage in distinct ways.
In the study conducted by Ely [39], re-
searchers examined the impact of adding lacto-
bacilli to fodder plants such as alfalfa, corn,
sorghum, and wheat. The treatment involved
using 5 grams of dry Lactobacillus acidophilus
per kilogram of fresh fodder, and the fermenta-
tion process was analyzed. The fermentation re-
sults showed no significant differences between
the inoculated and control groups, indicating
that the addition of bacterial additives did not
provide any benefits. The lack of success with
the bacterial additive in this study may be at-
tributed to several factors: 1) a readily available
energy source for microbes is essential for con-
trolled fermentation, and 2) the added microbes
must be able to compete with the existing epi-
phytic population for successful silage fermen-
tation. For high-quality silage from corn and al-
falfa fodder, both conditions must be met; oth-
erwise, the bacterial additive will not enhance
fermentation quality. In the case of wheat fod-
der, it appears that both conditions were lacking,
and even high doses of the bacterial additive
failed to lower the pH or prevent the develop-
ment of secondary harmful fermentation pro-
ducts. Additionally, Burghardi et al. (1980)
demonstrated that bacterial additives can be ad-
vantageous when the soluble carbohydrate con-
tent in the plant is low.
In the experiment of Adesogan et al. [1], the
effect of a bacterial additive containing Pedio-
coccus pentozeus in the amount of 105 and Lac-
tobacillus buccaneri in the amount of 105 was
used on bermuda grass fodder. pH was lower
throughout the fermentation period in the treat-
ment containing bacterial additive, and the re-
covery of dry matter from day 4 and soluble
carbohydrates from day 30 to the end of the pe-
riod was higher in the treatment containing bac-
terial additive. NDF was not affected, but ADF
was reduced in the additive treatment. The
amount of acetic acid, propionic acid, lactic
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26
acid, ethanol, lactate to acetate ratio, lactic acid
producing bacteria, yeasts and molds were not
affected by the additive after 60 days of ensi-
ling. The amount of butyric acid and total acids
produced in the bacterial additive treatment de-
creased and finally it was reported that access to
fermentable carbohydrates is more effective on
the rate of primary fermentation in tropical
plants with low soluble carbohydrate content
than desirable bacterial additives. But in gene-
ral, the effectiveness of the bacterial additive in
the final stages of fermentation confirms the
usefulness of the bacterial additive to overcome
homogeneous or non-homogeneous epiphytic
lactic acid producing bacteria.
Recent developments in creating dual-pur-
pose additives for both homogeneous and hete-
rogeneous lactic acid-producing bacteria have
been significant and yielded beneficial outco-
mes in this area. Adesogan and Salawu (2004)
found in their study that these dual-purpose bac-
terial additives enhanced silage fermentation but
did not influence aerobic stability. Conversely,
Filya et al. [43] demonstrated that these ad-
ditives improved both aerobic stability and
fermentation, differing from the findings of
Driehuis et al. [37]. This variation in results is
likely due to differences in the type of feed and
harmful organisms involved. Previous studies
have indicated that aerobic stability is mainly
linked to bacteria. McAllister et al. [80] pro-
posed that Lactobacillus buccaneri inhibits the
growth of aerobic yeasts in the initial phase, but
it seems unlikely to impact bacteria that damage
silage. In the experiment conducted by Kent et
al. [64], researchers examined the impact of a
bacterial additive that included Lactobacillus
plantarum and Pediococcus lactic acid on alfal-
fa silage. The findings indicated that the pH le-
vel dropped in the silage with the bacterial addi-
tive, while factors such as crude protein, ADF,
ADIN, and yeast count remained unchanged.
The swift generation of lactic acid in this study
likely contributed to the observed reduction in
pH in the treated silage.
In a meta-analysis conducted by Klein-
schmit and Kung [67], it was found that an addi-
tive containing Lactobacillus buccaneri at con-
centrations exceeding 105 CFU resulted in an
increase in pH, a decrease in lactic acid levels,
and a greater loss of nutrients from the surface
compared to the control group. Across all treat-
ments, the levels of acetic acid were lower than
what is typically observed in corn silage. How-
ever, starting from day 56, the levels of this or-
ganic acid were higher in the inoculated treat-
ments. Additionally, the ratio of lactate to ace-
tate in this study was greater than 3 in all sila-
ges, indicating a consistent fermentation of lac-
tic acid. In the experiment conducted by She-
perd and Combs [114], two types of additives
containing Lactobacillus plantarum and Pedio-
coccus cerevisiae, along with enzymes such as
cellulase, amylase, and pectinase, were applied
to first harvest alfalfa fodder. Throughout the
fermentation and ensiling process, the pH levels
were lower in the treatments that included both
types of bacterial and enzyme additives. The
levels of lactic acid and glucose remained con-
sistent across all treatments during the fermenta-
tion period, but by the end of the 177 days, the
control treatment had the lowest levels of both.
The production of acetate in the treatment with
the bacterial additive began to decline from the
8th day of ensiling and continued to decrease
until the end of the study. The experiment also
demonstrated that the bacterial additive could
lower the pH even when the dry matter content
limited fermentation. Based on his findings,
Jones proposed that incorporating bacteria into
alfalfa forage with high dry matter (over 35 %)
is more beneficial than adding fermentable sub-
strates [33].
In the experiment conducted by Bolsen
[19], a bacterial additive that included Lactoba-
cillus plantarum, Enterococcus faecium, dext-
rose, and a combination of the two was tested.
The bacterial additive alone did not affect the
pH, but the combination of the two significantly
lowered both pH and acetic acid levels, while
lactic acid levels increased. According to Spoel-
stra [118], alfalfa silage with bacterial additives
and fermentable substrates exhibited higher lac-
tic acid and lower acetic acid levels. The pre-
sence of the bacterial additive and dextrose led
to a decrease in ethanol levels. The microbio-
logical analysis of the silage revealed that the
populations of lactobacilli, pediococcus, leuco-
nostoc, enterobacter, yeast, mold, lactate-consu-
ming yeast, and clostridium remained unchan-
ged across treatments. The fermentation process
analysis indicated that the bacterial additive in
the fodder from the second and fourth harvests
resulted in the lowest pH on the first day, but
thereafter, a combination of lactic acid and dex-
trose-producing bacteria maintained the lowest
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27
pH. The lactic acid levels were highest on the
first day for both the bacterial additive alone
and the combination with dextrose, but from the
third day onward, only the treatment with both
bacterial additives and dextrose continued to
show this trend. Throughout the ensiling period,
the treatment with bacterial additives and dex-
trose had the lowest acetic acid levels from the
third day onward. In corn silage, no significant
differences were found between the control and
inoculated treatments regarding any fermenta-
tion indices.
Bolsen [19] found that alfalfa is an incon-
sistent forage and challenging to ensile. The ad-
dition of 2 % dextrose, a bacterial additive, or a
combination of both did not lead to successful
microbial activity for improved fermentation
during the ensiling process. However, these ad-
ditions significantly enhanced fermentation effi-
ciency, and the researchers ultimately achieved
the best fermentation results with a combination
of the two additives. Bolsen and his team con-
cluded that the lactic acid-producing epiphytic
population on the entire corn plant is substantial
and predominantly uniform. As a result, the bac-
teria from the bacterial additive were unable to
dominate during ensiling and did not demon-
strate their intended effects. Additionally, the
natural properties of the corn plant are condu-
cive to producing high-quality silage without
the need for any additives. In the study conduct-
ed by Aksu et al. [3], the impact of bacterial ad-
ditives, molasses, and formic acid was exami-
ned. The experiment utilized uniform lactic
acid bacteria, and it was found that the level of
lactic acid rose, while the levels of butyric acid
and acetic acid remained unchanged. In a study
conducted by Filya et al. [43], the effects of
Lactobacillus buccaneri, Lactobacillus planta-
rum, and a combination of both at a concentra-
tion of 106 CFU were examined on corn and
sorghum fodder with low dry matter content.
The results showed that after two days of ensi-
ling, the levels of acetic acid were higher in si-
lages treated with Lactobacillus buccaneri and
the combination of both additives compared to
other treatments. This trend continued on days
4, 8, and 15 post-ensiling. During the fermenta-
tion process, there was a decrease in pH and wa-
ter-soluble carbohydrates, while the concentra-
tions of lactic acid, acetic acid, ethanol, and
ammonia nitrogen increased. After 90 days of
ensiling, silages treated with Lactobacillus buc-
caneri exhibited a higher pH than those inocu-
lated with Lactobacillus plantarum, the combi-
nation of both, and the control group. Addition-
ally, silages treated with Lactobacillus planta-
rum and the combination of both Lactobacillus
strains had higher lactic acid levels compared to
the control silages treated solely with Lactoba-
cillus buchneri. The ammonia nitrogen levels in
silages treated with Lactobacillus plantarum
and the combination were lower than those in
silages treated with Lactobacillus buchneri
alone. Furthermore, the loss of dry matter in si-
lages treated with Lactobacillus buchneri was
greater compared to the control and those trea-
ted with both Lactobacillus strains. Lastly, the
control and silages treated with Lactobacillus
plantarum retained more residual water-soluble
carbohydrates than those treated with Lactoba-
cillus buchneri and the combination of both
strains.
In the experiment conducted by Ranjit and
Kung [110], researchers examined the impact of
an additive with Lactobacillus buchneri at two
different levels, along with two strains of Lac-
tobacillus plantarum and a treatment that in-
cluded a propionic acid buffer. The findings in-
dicated that there were no significant differences
in pH across the various treatments. In the
treatment with a high dose of Lactobacillus
buchneri, the lactate levels decreased while ace-
tate levels increased significantly. Additionally,
propionate levels rose in the treatment with the
propionate buffer. Butyrate levels could not be
detected in any of the treatments. The ratio of
lactate to acetate decreased in the high-dose
Lactobacillus buchneri treatment and one of the
Lactobacillus plantarum treatments, while it in-
creased significantly in the other Lactobacillus
plantarum treatment. Furthermore, the number
of yeasts diminished in the high-dose Lactoba-
cillus buchneri treatment. None of the Lacto-
bacillus plantarum strains achieved a uniform
fermentation in this study, suggesting that these
bacteria lacked the strength to outcompete the
existing epiphytic microbial population.
In a study by Filya et al. [44], researchers
examined the impact of 14 different bacterial
additives on alfalfa fodder during its first and
second cuts. All additives were applied at a con-
centration of 106 CFU per gram of DM. In the
first cut, all additives containing Enterococcus
faecium lowered the pH compared to the cont-
rol group, although two of the heterogeneous
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28
additives resulted in a smaller pH reduction than
anticipated. The levels of lactate increased by
2.5 % to 106 % in treatments with bacterial ad-
ditives, while seven additives raised the acetic
acid levels by 15 % to 255 %. Consequently, the
lactate-to-acetate ratio was lower than in the
control group for only three additives. Additio-
nally, two additives with Lactobacillus buchne-
ri and Enterococcus faecium produced higher
ethanol levels than the control. In the second
cut, only five additives significantly lowered
the pH, with Lactobacillus buchneri showing
the highest pH. Most additives, except for two,
increased lactic acid levels. Silaged alfalfa with
Lactobacillus buchneri and Lactobacillus pen-
tazeus had the highest acetate and ethanol con-
centrations, and these were the only treatments
with a low lactate-to-acetate ratio. After fermen-
tation, the control treatment in the first cut re-
tained more soluble carbohydrates, while the
lowest amounts were found in treatments with
Enterococcus faecium, Lactobacillus pentozeus,
and two strains of Lactobacillus buchneri. In the
second cut, the control treatment had intermedi-
ate levels, with the lowest values in treatments
containing Lactobacillus buchneri and the hig-
hest in those with Enterococcus faecium. Overall,
the concentrations of NDF, ADF, and acid-inso-
luble lignin were greater in both cuts compared
to non-ensiled forage, attributed to dry matter
loss from fermentation and respiration, which
led to sugar loss and an increased NDF ratio.
Ultimately, bacterial additives positively influ-
enced silage characteristics by lowering pH and
enhancing lactic acid production.
According to Ridwan et al. [112], incorpo-
rating lactic acid bacteria (LAB) significantly
improves the silage microbiome and quality by
changing the diversity of bacteria and the meta-
bolic byproducts of the silage materials, ensu-
ring safe preservation.
In the study conducted by Weinberg et al.
[130], researchers examined the impact of
10 different bacterial additives on corn and
wheat silage. For ensiled wheat, the pH levels
remained consistent across all treatments, while
the dry matter content was lower in six of the
additives compared to the control, but higher in
two additives that included Lactobacillus buch-
neri. The NDF content was elevated in four bac-
terial additives and decreased in two that con-
tained Lactobacillus buchneri. Lactic acid levels
were higher in treatments with Lactobacillus
plantarum and Enterococcus faecium, while
lower levels were found in treatments with Lac-
tobacillus plantarum, with no significant diffe-
rences in other treatments compared to the con-
trol. Ethanol levels increased in six of the bacte-
rial additives, with the highest concentration
linked to Pediococcus pentazeus and the lowest
in two treatments with Lactobacillus buchneri.
Acetic acid levels were highest in five additives
containing Enterococcus faecium and Lactoba-
cillus plantarum, and lowest in one treatment
with Lactobacillus pentazeus. Lactic acid-pro-
ducing bacteria were more prevalent than in
the control in eight treatments. In silage corn,
five treatments had a higher pH than the control,
and none of the additives were able to lower the
pH compared to the control. All treatments had
lower dry matter than the control, with the lo-
west amount associated with Lactobacillus
buchneri. NDF levels were higher in eight addi-
tives, with the highest observed in the Lactoba-
cillus buchneri treatment. Lactic acid levels
were higher in three treatments and lower in
three compared to the control, with the highest
from Lactobacillus pentozeus and the lowest
from Lactobacillus plantarum. Ethanol levels
were greater in all treatments compared to the
control, with the highest from Enterococcus
faecium. Acetic acid was elevated in eight addi-
tives, with the lowest in the Lactobacillus pen-
tazos treatment. Lactic acid-producing bacteria
were more abundant than in the control in all
treatments except for Lactobacillus buchneri.
Guo et al. [50] found that in different types of
silages, lactic acid bacteria (LAB) inoculants
influence the composition of microbial commu-
nities in various ways, depending on the exis-
ting microbiota on fresh forage. Essentially,
LAB inoculants streamline the relationships
between bacterial species to improve fermenta-
tion quality.
Na et al. [94] studied the effects of six
common commercial LAB additives (Lactoba-
cillus plantarum, L. buchneri, and Enterococcus
faecalis; L. plantarum and L. casei; L. planta-
rum and L. buchneri; L. plantarum, L. buchneri,
L. casei, and Pediococcus acidilactici; L. plan-
tarum; and L. buchneri, P. acidilactici) on the
bacterial community and fermentation quality of
alfalfa silage. They showed that using commer-
cial LAB additives during the ensiling of alfalfa
enhanced fermentation quality, aided in preser-
vation, and changed the bacterial composition
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29
of the final silage. In the Control silage, Lacto-
bacillus, Enterococcus, and Pediococcus were
the predominant bacteria. According to Kim et
al. [65], LAB inoculants enhance the quality of
silage and mitigate DM losses during prolonged
storage. LAB represents one of the most effec-
tive organic additives for the regulation of unde-
sirable bacterial proliferation in silage. These
inoculants facilitate the conversion of water-
soluble carbohydrates and complex secondary
metabolites present in forage crops into organic
and mineral acids, thereby effectively inhibiting
pathogenic growth and improving the nutritional
quality of silage for livestock.
Ling et al. [77] indicated that the incorpora-
tion of LB enhanced the relative abundance of
Lactobacillus and contributed to the improved
quality of alfalfa silage. The research conducted
by Jiang et al. [59] demonstrated that the incor-
poration of a compound comprising LAB and
sugar sources significantly enhanced the fer-
mentation quality, nutrient profile, and microbi-
al diversity of high-moisture alfalfa silage.
Conclusion
The goal of ensiling is to preserve fresh fo-
rage crops or other types of biomass for later
use. The quality of silage can be enhanced by
incorporating different bacterial inoculants,
which help during fermentation, storage, and
feeding by improving fermentation processes,
encouraging beneficial microbial diversity, and
inhibiting harmful microorganisms. Alfalfa is
the most important forage, and microbial addi-
tives can enhance its silage preparation.
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78010-2
Received 11.09.2024
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36
https://doi.org/10.35868/1997-3004.40.3-36
УДК 579.64:636.085
ВИКОРИСТАННЯ МІКРОБНИХ ДОБАВОК
ПІД ЧАС ПРИГОТУВАННЯ СИЛОСУ З ЛЮЦЕРНИ
(огляд літератури)
M. Бешараті1, M. Лакнер2
1Тебрізський університет, Іран
2Університет прикладних наук «Технікум Відня», Австрія
e-mail: maximilian.lackner@technikum-wien.at
Зелена маса люцерни є доволі проблемною для сінажування через підвищений вміст біл-
ка, низьку кількість водорозчинних вуглеводів, низький вміст сухої речовини та високу буфе-
рну здатність. У зв’язку з цим нещодавно було запропоновано нові підходи до покращення
виробництва сінажу за допомогою добавок, серед яких в останні роки активно використо-
вуються силосні закваски. Бактеріальні добавки використовуються для підвищення якості
силосу сільськогосподарських культур, з особливим акцентом на сінаж. Основною метою
додавання молочнокислих бактерій у сінаж є пригнічення проліферації небажаних мікроор-
ганізмів, включно з представниками Clostridium і Enterobacteriaceae. Це досягається швид-
ким підвищенням концентрації іонів водню до порогового значення, несприятливого для рос-
ту цих небажаних бактерій. Останні дослідження функцій бактеріальних добавок при сило-
суванні та сінажуванні біомаси сільськогосподарських культур свідчать про значний по-
тенціал для покращення продукту не лише як ферментованого корму, але й для доставки
пробіотичних мікроорганізмів, які можуть принести користь здоров’ю тварин. У статті
представлено комплексний огляд процесу приготування силосів та критично оцінено низку
досліджень щодо їхньої якості, а також впливу бактеріальних добавок на сінаж із люцерни.
Якість силосів можна підвищити шляхом додавання різних бактеріальних інокулянтів, які
допомагають під час бродіння, зберігання та годівлі, покращуючи процеси бродіння, стиму-
люючи розвиток різноманітних корисних бактерій та пригнічуючи проліферацію шкідливих
мікроорганізмів. Люцерна є одним із найважливіших кормів, і мікробні добавки можуть по-
кращити приготування сінажу економічно ефективним і екологічно прийнятним способом.
Ключові слова: люцерна, бактеріальна добавка, буферна ємність, сінаж, в’ялення.
Отримано 11.09.2024
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 40.
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| id | oai:ojs2.smic.in.ua:article-529 |
| institution | Agriciltural microbiology |
| keywords_txt_mv | keywords |
| language | English |
| last_indexed | 2026-07-23T01:16:11Z |
| publishDate | 2024 |
| publisher | Institute of Agrocultural Microbiology and Agro-industrial Manufacture of NAAS of Ukraine |
| record_format | ojs |
| resource_txt_mv | smicinua/38/74f2660f7a4785083cf5f15e2505b638.pdf |
| spelling | oai:ojs2.smic.in.ua:article-5292026-07-22T10:10:52Z USE OF MICROBIAL ADDITIVES IN ALFALFA SILAGE PREPARATION (a review) ВИКОРИСТАННЯ МІКРОБНИХ ДОБАВОК ПІД ЧАС ПРИГОТУВАННЯ СИЛОСУ З ЛЮЦЕРНИ (огляд літератури) Бешараті, M. Лакнер, M. alfalfa, bacterial additive, buffer capacity, silage, wilting люцерна, бактеріальна добавка, буферна ємність, сінаж, в’ялення Alfalfa poses challenges for ensiling because of its elevated protein levels, low amounts of water-soluble carbohydrates, low dry matter content, and high buffering capacity. As a result, there has been a recent push to improve silage production using additives. In recent years, silage additives have been employed to enhance the quality of alfalfa silage. Bacterial additives are employed to enhance the quality of crop silage, with a particular emphasis on hay silage. A primary objective of incorporating lactic acid bacteria into silage is to inhibit the proliferation of undesirable microorganisms, including Clostridium and Enterobacteriaceae. This is achieved by swiftly elevating the hydrogen ion concentration to a threshold that is inhospitable for the growth of these detrimental bacteria. Recent insights into the functions of bacterial additives in crop silage suggest significant potential for enhancing silage, not just as a fermented feed, but also to deliver probiotic substances that can benefit animal health. This article provides a comprehensive overview of the silage preparation process and critically assesses a range of studies concerning the quality of silage, as well as the impact of bacterial additives on alfalfa silage. The quality of silage can be enhanced by incorporating different bacterial inoculants, which help during fermentation, storage, and feeding by improving fermentation processes, encouraging beneficial microbial diversity, and inhibiting harmful microorganisms. Alfalfa is the most important forage, and microbial additives can enhance its silage preparation in a cost-effective and environmentally friendly way. Зелена маса люцерни є доволі проблемною для сінажування через підвищений вміст білка, низьку кількість водорозчинних вуглеводів, низький вміст сухої речовини та високу буферну здатність. У зв’язку з цим нещодавно було запропоновано нові підходи до покращення виробництва сінажу за допомогою добавок, серед яких в останні роки активно використовуються силосні закваски. Бактеріальні добавки використовуються для підвищення якості силосу сільськогосподарських культур, з особливим акцентом на сінаж. Основною метою додавання молочнокислих бактерій у сінаж є пригнічення проліферації небажаних мікроорганізмів, включно з представниками Clostridium і Enterobacteriaceae. Це досягається швидким підвищенням концентрації іонів водню до порогового значення, несприятливого для росту цих небажаних бактерій. Останні дослідження функцій бактеріальних добавок при силосуванні та сінажуванні біомаси сільськогосподарських культур свідчать про значний потенціал для покращення продукту не лише як ферментованого корму, але й для доставки пробіотичних мікроорганізмів, які можуть принести користь здоров’ю тварин. У статті представлено комплексний огляд процесу приготування силосів та критично оцінено низку досліджень щодо їхньої якості, а також впливу бактеріальних добавок на сінаж із люцерни. Якість силосів можна підвищити шляхом додавання різних бактеріальних інокулянтів, які допомагають під час бродіння, зберігання та годівлі, покращуючи процеси бродіння, стимулюючи розвиток різноманітних корисних бактерій та пригнічуючи проліферацію шкідливих мікроорганізмів. Люцерна є одним із найважливіших кормів, і мікробні добавки можуть покращити приготування сінажу економічно ефективним і екологічно прийнятним способом. Institute of Agrocultural Microbiology and Agro-industrial Manufacture of NAAS of Ukraine 2024-10-23 Article Article Рецензована Стаття application/pdf https://smic.in.ua/index.php/journal/article/view/529 10.35868/1997-3004.40.3-36 Agricultural microbiology; Vol. 40 (2024): Agriciltural microbiology; 3-36 Сільськогосподарська мікробіологія; Том 40 (2024): Сільськогосподарська мікробіологія; 3-36 1997-3004 10.35868/1997-3004.40 en https://smic.in.ua/index.php/journal/article/view/529/598 Авторське право (c) 2024 M. Besharati, M. Lackner https://creativecommons.org/licenses/by/4.0 |
| spellingShingle | люцерна бактеріальна добавка буферна ємність сінаж в’ялення Бешараті, M. Лакнер, M. ВИКОРИСТАННЯ МІКРОБНИХ ДОБАВОК ПІД ЧАС ПРИГОТУВАННЯ СИЛОСУ З ЛЮЦЕРНИ (огляд літератури) |
| title | ВИКОРИСТАННЯ МІКРОБНИХ ДОБАВОК ПІД ЧАС ПРИГОТУВАННЯ СИЛОСУ З ЛЮЦЕРНИ (огляд літератури) |
| title_alt | USE OF MICROBIAL ADDITIVES IN ALFALFA SILAGE PREPARATION (a review) |
| title_full | ВИКОРИСТАННЯ МІКРОБНИХ ДОБАВОК ПІД ЧАС ПРИГОТУВАННЯ СИЛОСУ З ЛЮЦЕРНИ (огляд літератури) |
| title_fullStr | ВИКОРИСТАННЯ МІКРОБНИХ ДОБАВОК ПІД ЧАС ПРИГОТУВАННЯ СИЛОСУ З ЛЮЦЕРНИ (огляд літератури) |
| title_full_unstemmed | ВИКОРИСТАННЯ МІКРОБНИХ ДОБАВОК ПІД ЧАС ПРИГОТУВАННЯ СИЛОСУ З ЛЮЦЕРНИ (огляд літератури) |
| title_short | ВИКОРИСТАННЯ МІКРОБНИХ ДОБАВОК ПІД ЧАС ПРИГОТУВАННЯ СИЛОСУ З ЛЮЦЕРНИ (огляд літератури) |
| title_sort | використання мікробних добавок під час приготування силосу з люцерни (огляд літератури) |
| topic | люцерна бактеріальна добавка буферна ємність сінаж в’ялення |
| topic_facet | alfalfa bacterial additive buffer capacity silage wilting люцерна бактеріальна добавка буферна ємність сінаж в’ялення |
| url | https://smic.in.ua/index.php/journal/article/view/529 |
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