animals

Review
Probiotics and Postbiotics as Substitutes of Antibiotics in Farm
Animals: A Review
Daria Zamojska 1,2, * , Adriana Nowak 2, * , Ireneusz Nowak 3 and Ewa Macierzyńska-Piotrowska 1

1 Polwet-Centrowet Sp. z o.o., M. Konopnickiej 21, 98-100 Lask, Poland; [email protected]

2 Department of Environmental Biotechnology, Lodz University of Technology, Wolczanska 171/173,
90-530 Lodz, Poland
3 Faculty of Law and Administration, University of Lodz, Kopcinskiego 8/12, 90-232 Lodz, Poland;
[email protected]
* Correspondence: [email protected] (D.Z.); [email protected] (A.N.)

Simple Summary: Breeders are searching for methods to protect farming animals against diseases
caused by pathogenic bacteria. The easiest way to fight bacteria is to use antibiotics. Unfortunately,
their abuse results in the presence of bacteria resistant to the most commonly used antibiotics in the
environment. The restrictions on the use of antibiotics have forced the search for natural and safe
ways to protect animals. It has been shown that the use of probiotics based on lactic acid bacteria
may have a positive effect on the growth and use of feed by broilers, on the stabilization of the
intestinal microbiota of chickens and pigs, and in the prevention of mastitis in dairy cows. The use of
probiotics (live, nonpathogenic microorganisms) and postbiotics (inanimate bacteria, cell components
or post-fermentation by-products) reduces the occurrence of pathogens in large-scale farms.






Abstract: Since 2006, the use of growth-promoting antibiotics has been banned throughout the
Citation: Zamojska, D.; Nowak, A.;
European Union. To meet the expectations of livestock farmers, various studies have been carried
Nowak, I.; Macierzyńska-Piotrowska,
out with the use of lactic acid bacteria. Scientists are trying to obtain the antimicrobial effect against
E. Probiotics and Postbiotics as
Substitutes of Antibiotics in Farm
the most common pathogens in large-scale farms. Supplementing the diet of broilers with probiotics
Animals: A Review. Animals 2021, 11, (live, nonpathogenic microorganisms) stabilized the intestinal microbiota, which improved the
3431. https://doi.org/10.3390/ results of body weight gain (BWG) and feed intake (FI). The positive effect of probiotics based on
ani11123431 lactic acid bacteria has been shown to prevent the occurrence of diarrhea during piglet weaning.
The antagonistic activity of postbiotics (inanimate bacteria, cell components, or post-fermentation
Academic Editor: Valentina by-products) from post-culture media after lactobacilli cultures has been proven on Staphylococcus
Virginia Ebani aureus—the pathogen most often responsible for causing mastitis among dairy cows. The article aims
to present the latest research examining the antagonistic effect of lactic acid bacteria on the most
Received: 20 October 2021
common pathogens in broilers, piglets, pigs, and cow farms.
Accepted: 29 November 2021
Published: 1 December 2021
Keywords: broiler; piglet; pig; bovine; lactic acid bacteria; probiotic; postbiotic; antibiotic; farm animal

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1. Introduction
iations.
The invention of antibiotics is considered to be one of the greatest discoveries of the
twentieth century and made it possible to control many diseases, but with their “over-
normative” use, new threats appeared, including antibiotic resistanceto both human and
Copyright: © 2021 by the authors.
veterinary medicinal products for animals [1]. It is obvious that the consumption of meat
Licensee MDPI, Basel, Switzerland.
and meat products from animals fed with antibiotics (including their residues) is undoubt-
This article is an open access article
edly harmful to the health of the latter, and even more so to people, because even in a low
distributed under the terms and concentration and regardless of the period time, it is not physiologically indifferent [2–5]. In
conditions of the Creative Commons other words, in animal husbandry, antibiotics should only be used for therapeutic purposes
Attribution (CC BY) license (https:// and not, inter alia, for reducing falls due to the crowding of animals in a small area, im-
creativecommons.org/licenses/by/ proving their condition, or stimulating growth and development. Thus, it is unacceptable
4.0/). to legally legalize the maximization of the fattening of farm animals through the use of

Animals 2021, 11, 3431. https://doi.org/10.3390/ani11123431 https://www.mdpi.com/journal/animals

Animals 2021, 11, 3431 2 of 20

antibiotics solely for strictly economic reasons [5]. The development of agriculture has
led to the introduction of intensive livestock to satisfy parts of society around the world.
Unfortunately, intensive industrial breeding increases density on farms and promotes the
development of various diseases [6,7]. Crowded livestock production conditions facilitate
the transmission of zoonotic pathogens, such as parasites, fungi, viruses, and bacteria
among individuals [8,9].
In 2019, the EU produced 43.5 million tons of meat, which was around 10% less than in
2018. From the number, approximately 52% came from pigs (22.8 million tons), 6.9 million
tons were bovine meat, and 13.3 million tons were poultry. In the same year, the EU
produced 158.2 million tons of raw milk. The European Union bred 143 million pigs, and
77 million cattle were also bred. The main producer of poultry meat in the EU was Poland
(2.6 million tons) [10].
The sales of pharmaceutical forms of drugs in 31 countries accounted for 87.7% of
the total sales of veterinary products: premixes accounted for 26.9%, oral powder for 9%,
and solutions for 51.8%. The sales of intramammary preparations amounted to 0.6%. In
the years 2011–2018, a decrease in sales by 34.6% was observed in 25 countries. Sales of
tetracyclines (30.7%), penicillins (28.8%), and sulfonamides (8.4%) in mg/PCU accounted
for 67.9% of total sales in 2018 [11]. One experiment showed an increased productivity of
chickens after the administration of subtle doses of antibiotics, such as chlorotetracycline,
virginiamycin, and amoxicillin. In animals fed with antibiotics, the feed conversion ratio
(FCR) decreased, but in the feces samples, heterotrophic bacteria with antibiotic resistance
were detected [12]. The overuse of antimicrobial agents in animal husbandry causes more
and more frequent occurrences of pathogenic bacteria resistant to popular antibiotics [13].
In the review, the positive effect of the use of probiotics on achieving the balance
of intestinal microorganisms and the antibiotic-like effect in inhibiting the growth of
pathogenic microorganisms found in slaughter farms in chickens, pigs, piglets, and cattle is
described. According to the definition used by The International Scientific Association for
Probiotics and Prebiotics (ISAPP), a probiotic is a live, non-pathogenic microorganism with
a positive effect on the host [14]. According to Williams (2010), microorganisms classified as
probiotics are most often lactic acid bacteria (LAB), but also Bacillus cereus and Enterococcus
faecalis [15]. Probiotic bacteria, in particular LAB, are used to obtain fermented dairy
products, e.g., Greek yogurt (S. thermophilus ACA-DC 26), feta cheese (L. plantarum ACA-
DC 2640) [16], kefir [17,18], and fermented vegetables, e.g., sauerkraut [19,20]. Consuming
probiotics shows a health-promoting effect for humans [21,22]. Likewise, probiotics protect
food against the development of pathogens and spoilage [22,23]. The review also describes
the positive effects of postbiotics on the animals’ gut microbiome.
The latest ISAPP definition says that postbiotics are inanimate microorganisms or
bacterial cell components and that post-fermentation by-products such as lactic acid and
short chain fatty acids (SCFA) positively affect the host [14]. According to Pandey et al.
(2015), a synbiotic is a food component or dietary supplement consisting of an appropriate
combination of a probiotic and a prebiotic [24]. The ISAPP gave a more precise definition
of synbiotic. Synbiotic causes a positive effect on the health of the host, and it is a mixture
of microorganisms and nutrients used by the host’s microorganisms [25]. A prebiotic is
a substance used by microorganisms that has a positive effect on its growth [14]. In the
cited example, the prebiotic was the polysaccharide inulin [26] added to the probiotic mix
of LAB bacteria [27] (Figure 1).
This article discusses the latest research on the effects and benefits of LAB and their
metabolites on the intestinal microbiota of the most common farm animals: poultry, bovines,
and pigs. The review covers the newest literature on the application of probiotics and
postbiotics in nutrition and in improving the welfare of farm animals. In addition, the
future legal regulations regarding antibiotics in the European Union are discussed.
Animals 2021, 11, 3431 3 of 20

Figure 1. Effect of antibiotics application in animal husbandry.

2. The Welfare Improving Poultry Farming

The most common pathogen in the poultry gastrointestinal tract in broiler breeding is
the Salmonella species. Due to the limitation of the use of antibiotics, research is conducted
with the use LAB bacteria in order to limit the colonization of the intestines by Salmonella
sp. Wang et al. vaccinated hatched chicks with a strain of Lactiplantibacillus plantarum
LTC-113. Studies have shown protection against Salmonella Typhimurium by reducing
gut colonization and stabilizing tight junction gene expression in intestinal epithelial cells
among treated chickens. In the control group, a Salmonella infection disrupted the intestinal
epithelial barrier [28].
In contrast, another study reported that orally administered Lactobacillus johnsonii
reduced intestinal colonization by Salmonella and Clostridium perfringens [29]. In another
variant, the probiotic was given combined with two strains: Ligilactobacillus salivarius
and Enterococcus faecium. The probiotic reduced intestinal colonization by Salmonella
Enteritidis and did not cause weight loss or damage to the gastrointestinal mucosa [30].
In another experiment, the use of the E. faecium probiotic in feed increased egg weight,
serum FSH (follicle-stimulating hormone) levels, and decreased Bacteroidetes (phylum) in
low reproductive individuals [31]. Salmonella enters the bloodstream and then goes to the
liver and spleen through damage to the intestinal barrier, which has been confirmed in
many studies [32–34].
The administration of LAB bacterial cell membrane extracts to chickens reduced the
Salmonella Enteritidis infection. Considerable protection of the intestinal epithelium against
the effects of infection was evident [32]. The next experiment was very similar, however, a
commercial probiotic based on LAB with proven efficacy against S. Enteritidis was used for
treatment. The study aimed to understand the effect of a probiotic on intestinal colonization
and intestinal permeability in infected chickens. The results were very promising. After
infection of the control group with Salmonella pathogens, heterophilia and lymphopenia
were observed as well as an increase in basophils and eosinophils compared to the chickens
treated with a probiotic based on LAB bacteria. In the control group, increased intestinal
permeability was also found [33]. In another experiment, the probiotic Bacillus subtilis

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Animals 2021, 11, 3431 4 of 20

C-3102 was used to control S. enteric serovar enteritidis LM-7. The specific pathogen-free
chicks became infected by administering an appropriate dose of Salmonella in the food.
Supplementation with B. subtilis feed reduced Salmonella infections and may accelerate the
clearance of pathogens in the liver, cecum, and spleen of chicken farms [34].
Han et al. (2017) orally administered Pediococcus acidilactici mutants to chickens, mod-
ulating the microbiota and reducing the number of Salmonella enterica serovar Gallinarum,
which is often found in the poultry industry. The study proved the antimicrobial activity of
P. acidilactici. After treatment of the cultures with proteinase, the antimicrobial activity de-
creased, which may suggest the production of proteinaceous substances such as bacteriocin
by P. acidilactici [35].
Another common pathogen on farms is Campylobacter sp. The pathogen is rapidly
transmitted on poultry farms via the fecal–oral route. The study of Ščerbová et al. (2016)
assessed the inhibition spectrum of various enterocins against Campylobacter sp. Ente-
rocins are protein substances with antibacterial activity metabolized mainly by enterococci.
The isolated strains from poultry farming were divided into two groups, Campylobacter
jejunum, and Campylobacter coli. Interestingly, the strains showing resistance to antibiotics
displayed sensitivity to at least one of the nine eneterocins tested [36]. On the other hand,
Razmyar et al. (2017) showed that C. perfringens secretes bacteriocins, which may be re-
sponsible for facilitating intestinal colonization and causing intestinal inflammation by this
pathogen [37]. Supernatants from Lactobacillus acidophilus NCFM, Lactobacillus crispatus
JCM 5810, Lactobacillus gallinarum ATCC 33199, and Lactobacillus helveticus CNRZ32 cultures
inhibited the growth of C. jejuni in vitro. After the analyses, the substance responsible for
limiting the growth of C. jejuni was partly lactic acid. Subsequently, in in vivo studies, LAB
was administered to broilers on the day of hatching, on the fourth day after hatching, and
14 days after hatching, challenged with C. jejuni F38011. Each of the four strains limited
the colonization of the pathogen. It was most effective in limiting the colonization of
L. crispatus JCM 5810 [38].
Another pathogenic bacterium on chicken farms is avian E. coli (APEC). Birds infected
with APEC show macroscopic changes in air sacs and lungs [39]. In the experiment
of Li et al. (2021), Lactobacillus animalis (ATCC 35046), Lactobacillus reuteri (ATCC 2837),
and Lactobacillus rhamnosus (ATCC 23272) strains were injected into the eggs to inhibit
APEC infection. However, no differences were noted in APEC-like strains infection in the
probiotic-administered group and control group [40]. This result was completely different
from that obtained in the experiment in which the probiotic mix with B. subtilis, Clostridium
butyricum, and L. plantarum was used. The applied probiotic lowered the E. coli index in
infected chickens. The positive effect of the probiotic on the modulation of the intestinal
microbiota of broilers has also been proven [41].
The protection of poultry farms against microbes is one of the most important factors
in good breeding. However, it is also an important factor to improve European indicators
for weight gain and feed consumption. The study of Kierończyk et al. (2017) aimed to test
the effect of nisin on growth efficiency, morphological parameters, the activity of digestive
enzymes, the digestibility of nutrients, and the effect on intestinal morphology in chickens.
It was noticed that supplementing the diet of chickens with nisin, which is a bacteriocin
used in the preservation of food products [42], improved the body weight gain (BWG),
feed conversion ratio (FCR), and feed intake (FI) indexes [43]. According to Hsu et al.
(2004), nisin is a cyclic polypeptide that contains 34 amino acids. The lactic acid bacteria of
the genus Lactococcus lactis, carries out the fermentation process and, in addition to lactic
acid, produces the bacteriocin nisin. Nisin is a natural antibiotic against Gram-positive
bacteria [44,45]. Research has indicated that nisin can be used as a growth simulator without
adversely affecting the bird’s metabolism or immunity levels [28]. According to Kierończyk
et al. (2020), nisin can be considered as a new and natural growth promoter. It improves
digestibility and feeds conversion. By limiting the multiplication of pathogenic bacteria,
nisin has a positive influence on the modulation of the intestinal microbiota. In terms
Animals 2021, 11, 3431 5 of 20

of its antibacterial properties, it is similar to monensin, an antibiotic of the coccidiostatic

type [46].
The intestinal microbiota was modulated not only with probiotics but also with plant
feed additives. Wang et al. (2021) showed a positive effect of dietary purslane in the
experiment. Purslane (Portulaca oleracea L.) is an edible wild vegetable with medicinal
properties. The use of purslane in the feed increased the level of Lactobacillus and lowered
Escherichia/Shigella in the digestive tract of broilers. The growth of beneficial bacteria in
chicken intestines may promote high body weight gain [47]. Another study by Liang
et al. (2021) applied traditional Chinese medicine based on medicinal plants combined
with probiotics—a mix of B. subtilis and L. acidophilus. In the treatment of E. coli-infected
chickens, the mix inhibited the survival level of E. coli, reduced the rates of diarrhea and
mortality, improved body weight gain, and relieving pathological changes in the intestines
and liver were observed [48].
In another study confirming the beneficial effect of LAB on the intestinal microbiota
of fattening chickens conducted by Śliżewska et al. (2020), the effects of three variants of
synbiotics were compared with two variants of a commercial probiotic on the chicken’s
performance. The synbiotics were three combinations containing the following strains:
L. plantarum ŁOCK 0860, L. reuteri ŁOCK 1092, L. pentosus ŁOCK 1094, Saccharomyces
cerevisiae ŁOCK 0119, L. rhamnosus ŁOCK 1087, L. paracasei ŁOCK 1091, and 2% inulin
(prebiotic). Commercial probiotics included BioPlus YC (Bacillus licheniformis DSM 5749, B.
subtilis DSM 5750) and Cylactin (Enterococcus faecium NCIMB 10415). The positive effect of
synbiotics on the performance of fattening chickens and the balance of the intestinal micro-
biota was demonstrated. The number of beneficial microorganisms such as Bifidobacterium
sp. and Lactobacillus sp. increased in the intestines, and the number of pathogenic bacteria
such as Clostridium sp. and E. coli in the intestines and animal excretions decreased. The
change in the gut microbiome increased the levels of lactic acid and SCFA (short chain
fatty acid). This is another study confirming the beneficial effect of LAB on the intestinal
microbiota of fattening chickens [27].
The Gram-negative bacteria Gallibacterium anatis is responsible for the decrease in
the number of eggs laid by causing infections of the genital tract of hens and contributes
to increased mortality [49,50]. A study conducted by Zhang et al. (2021) showed the
antagonistic activity of the supernatant after the culture of Leuconostoc mesenteroides QZ1178
(a species of lactic acid bacteria) was used against G. anatis strains in vitro. The antag-
onistic effect was decreased upon increasing the pH. After analysis of the supernatant,
L. mesenteroides QZ1178 mainly produced lactic acid (29 mg/mL) and acetic acid (7 mg/mL),
which are probably responsible for its antibacterial properties [51]. The above information
is summarized in Table 1.

Table 1. Overview of the application of beneficial bacteria in poultry farming in an in vivo study.

Probiotic/Postbiotic/Synbiotic Form/Way of Administration Effect References

Lactiplantibacillus plantarum Protection against Salmonella Typhimurium;
vaccination [28]
LTC-113 stabilizing intestinal epithelial barrier
Reduction of Salmonella sofia and Clostridium
Lactobacillus johnsonii per os in feed [29]
perfringens
Ligilactobacillus salivarius 59 and
Decreased colonization by Salmonella Enteritidis
Enterococcus faecium per os in feed [30]
S1400
PXN33/probiotic
Increased egg weight, serum FSH * levels,
E. faecium/probiotic per os in feed [31]
decreased Bacteroidetes
Reduction of Salmonella eteric serovar enteritidis
Bacillus subtilis C-3102/probiotic per os in feed [34]
LM-7
Reduction of Salmonella enterica serovar
Pediococcus acidilactici per os in feed [35]
Gallinarum
Animals 2021, 11, 3431 6 of 20

Table 1. Cont.

Probiotic/Postbiotic/Synbiotic Form/Way of Administration Effect References

Lactobacillus acidophilus NCFM,
Lactobacillus crispatus JCM 5810,
Lactobacillus gallinarum ATCC per os in feed Inhibition of the growth of Campylobacter jejuni [38]
33199, and Lactobacillus helveticus
CNRZ32
B. subtilis MORI 91, Clostridium
Lowered rate of E. coli; positive modulation of the
butyricum M7 and L. plantarum per os in feed [41]
intestinal microbiota
K34/commercial probiotic mix
Natural growth promotor; positive influence on
Nisin/postbiotic per os in feed the modulation of the intestinal microbiota; [42,46]
limitation of pathogens in the gut
Chinese medicinal plants and B. Inhibition of E. coli; reduction of mortality;
per os in water [48]
subtilis, L. acidophilus (probiotic) improvement BWG*
L. plantarum ŁOCK 0860, L. reuteri
ŁOCK 1092, L. pentosus ŁOCK
1094, Saccharomyces cerevisiae Increase of Bifidobacterium sp. and Lactobacillus sp.;
per os in feed [27]
ŁOCK 0119, L. rhamnosus ŁOCK decrease in the level of Clostridium sp. and E. coli
1087, L. paracasei ŁOCK 1091 and
2% inulin (synbiotic)
The positive effect of intestinal health and
L. plantarum CCTCC M2016259,
intestinal microbiota, improved body weight and
Paenibacillus polymyxa per os in feed [52,53]
feed conversion in C. perfringens-infected;
CGMCC1.1711
increased intestinal SCFAs * levels
Increased body weight; reduced mortality;
L. acidophilus per os in feed improvement of the immune response in E. coli [54]
O157-challenged chickens
Increased FCR*, villus height to crypt depth ratio,
and number of Blautia, Faecalibacterium,
B. subtilis per os in feed Flavonifractor, Hydrogenoanaerobacterium, and [55]
Romboutsia; decreased Odoribacter; improvement
intestinal microbial composition
Increased calcium and phosphorus in plasma;
B. subtilis PB6 per os in feed increased bone mass and meat quality; [56]
improvement production and welfare
Increased level of Lactobacillus spp.; decreased
B. subtilis QST713 per os in feed level of E. coli and Enterococcus spp.; elongated villi; [57]
fewer deep crypts
Increased FCR*, numbers of goblet cells, and
superoxide dismutase activities in the jejunal [58]
mucosa; elongated villi
B. subtilis DSM 29784 per os in feed Improved health, weight, the tight junction
complex in necrotic enteritis-challenged broilers;
increased numbers of Butyricicoccus and [59]
Faecalibacterium in the intestine; raised expression
of INF-γ and IL-12
Inhibition of the growth of Salmonella typhimurium;
elongated villi; reduced the length of jejunum and
E. faecium PNC01 per os in feed [60]
ileum; increased number of Firmicutes and
Lactobacillus; reduced the number of Bacteroides
* FSH—follicle-stimulating hormone; BWG—body weight gain; SCFA—short-chain fatty acids; FCR—feed conversion ratio.

3. Prevention of the Effects of Piglet Weaning Based on the Use of LAB

The critical moment in breeding is weaning the piglets from sow on day 28. Pigs
are very sensitive to changes in their living environment. This is a very stressful time for
piglets, causing destabilization of the intestinal microbiota. During this time, digestive
disorders, diarrhea, growth retardation, and increased mortality occur [61–64]. The most
common pathogens affecting intestinal disorders and damage to intestinal villi are E. coli,
C. perfringens, Salmonella Choleraesuis, and Salmonella Typhimurium. In the case of infection,
Animals 2021, 11, 3431 7 of 20

the permeability of fluids to the intestinal lumen increases and diarrhea develops. There is
also an increase in pH, which prevents the multiplication of LAB [65].
Antibiotics are used to improve the intestinal microbiota and reduce the occurrence
of diarrhea caused by weaning. The overuse of antibiotics has resulted in the emergence
of pathogenic bacteria resistant to the basic antibiotics used in farming. Moreover, in the
case of pig farms, research on the use of LAB to improve the intestinal microbiota was
started [66].
Verso et al. (2018) isolated 595 pure cultures of bacteria from the small and large
intestines from pre and post-weaned piglets. The selected bacteria were antagonistics to
pathogens and were capable of producing antimicrobial compounds. First, the activity
against pathogens E. coli MC4100, S. Choleraesuis ATCC 29628, and Listeria innocua HPB13
was tested using the Double-Agar-Layer Technique Method. At total of 51.1% of all
strains showed antimicrobial activity and passed the next test to investigate the production
of antimicrobial compounds. For this, the pH of the supernatant was neutralized with
1M NaOH. Activity against previous pathogens and Staphylococcus aureus ATCC 6538,
Enterococcus faecalis ATCC 27275, Listeria monocytogenes LSD530, and S. Enterica ATCC 8387
was tested by the Agar-Well Diffusion Method. Loss of activity of some supernatants after
protease treatment may indicate the presence of bacteriocin-like substances. These studies
confirmed the ability of some LAB strains of the species L. salivarius and Lactobacillus
delbrueckii subsp. lactis isolated from the digestive tract of pigs to inhibit the growth of
potential pathogens by the production of organic acids in combination with bacteriocin-
like proteins [67]. In another experiment, piglets were fed with an immunobiotic feed
based on okara fermented soy milk with L. delbrueckii subsp. delbrueckii TUA4408L. The
beneficial intestinal microbiota improved, the amount of Lactobacillus and Lactococcus
increased, and the immunity also increased. The piglets showed better meat quality and
growth performance [68]. In addition, the use of L. delebureckii CCTCC M 207,040 as a
diet supplement by Chen et al. (2021) improved the gut structure resulting in increased
gut integrity in lipolisaccharides (LPS)-challenged piglets. LPS stress induced an increase
in the depth of the crypts in the jejunum and ileum. However, the use of L. delebureckii
dietary supplementation reduced crypt depth compared to the non-challenged controls
and LPS-challenged. Moreover, the TLRs-Btk-Nrf2 signaling pathway, which mediates
oxidative stress, was mitigated [69]. Similar results were obtained in weaned piglets with
C. butyricum ZJU-F1 and B. licheniformis. The intestinal permeability was reduced, and
the digestibility of nutrients and the expression of antimicrobial peptides in the ileum
improved [70]. In another experiment, Sobrino et al. (2021) isolated L. salivarius MP100
from sow’s milk and gave inoculated feed to pregnant sows and piglets. MP100 showed
antagonistic activity against the indicator bacteria: C. perfringens MP34, E. faecalis MP42,
S. aureus MP83, Streptococcus suis MP205, Trueperella pyogenes MP214, E. coli MP73 (F4)
and MP77 (F18), S. Typhimurium MP55, and Klebsiella pneumoniae MP87. The use of a
potential probiotic resulted in a microbiological and biochemical improvement in the gut
environment [71].
The Lactobacillus gasseri LA39 and Limosilactobacillus frumenti strains produce the sub-
stance gassericin A, which is a bacteriocin. A characteristic feature of this protein is that it
binds to the intestinal epithelium of the host and makes it resistant to diarrhea in weaned
piglets. Gassericin A bound to Keratin 19 in the plasma membrane of the intestinal epithe-
lium increased the absorption of fluid from the intestine and reduced its secretion. The early
weaning of piglets aims to shorten the slaughter cycle and improve the reproduction of the
sows. In the experiment, fecal microbiota were taken from healthy Congjiang miniature
pigs (a Chinese native pig breed) and administered orally to a commercial Landrace ×
Yorkshire (LY) pig, which shows frequent diarrhea after weaning. This treatment made
the LY immune to stress-related diarrhea at weaning. The L. gasseri LA39 and L. frumenti
strains can be an alternative to antibiotics in the prevention of diarrhea during increased
stress in piglets [72]. In one study, the administration of L. salivarius (strains 144 and
160) to suckling piglets early in life resulted in an increase of the amount of Lactobacillus
Animals 2021, 11, 3431 8 of 20

in the gastrointestinal tract and a reduction in the number of Bacteroides and Fibrobac-
ter. The incidence of diarrhea during the most stressful time of weaning also decreased.
The conclusions from the conducted research indicated the use of supplementation with
L. salivarius 144 isolated from healthy pigs with a high BMI (body mass index), as it had
a beneficial effect on increasing the height of intestinal villi, which influenced the pigs’
growth efficiency. The same strain showed a reduction in the amount of Clostridium sp. in
the feces [73].
Another way of administering LAB strains to pigs was to use of microcapsulation,
i.e., administering LAB strains in sealed gelatin–alginate capsules. This method protects
microorganisms against unfavorable conditions in the digestive tract. In the study of Le
et al. (2019), the strains were isolated from traditional Vietnamese fermented yogurt. After
the bacteria had multiplied, they were centrifuged, the supernatant was removed, and the
remaining biomass was encapsulated. The L. plantarum SC01 strain showed antagonistic
activity against E. coli, S. aureus, B. subtilis, Salmonella sp., and L. monocytogenes. It has been
shown that the concentration of 2.5% (w/v) of alginate and 6% (w/v) of gelatin increases
the production of a highly active compound that inhibits pathogens by LAB [74]. On the
other hand, Pupa et al. (2021) used spray drying microencapsulation of L. plantarum 22F.
The application of this method as well as alginate and chitosan usage for the production
of capsules extended the viability of probiotic bacteria. The increased performance of
pigs after microencapsulated probiotic supplementation was comparable to the use of live
bacterial cultures. The alive bacteria: L. plantarum (strains 22F and 25F) and P. acidilactici
(strain 72N) were administered as potential probiotic supplements. The best effect in
reducing pathogenic intestinal strains (Enterobacteriaceae) and modulating lactobacilli in the
intestinal tract was obtained with the use of P. acidilactici 72N. The administration of the
probiotics to young animals resulted in the improvement of intestinal integrity, elongation
of intestinal villi in the jejunum, the appearance of microorganisms positively influencing
the intestinal microbiome, and improved growth of individuals in the rearing cycle [75].
The weakening of the animals also occurs during pregnancy and the lactation of
sows. Wang et al. (2014) isolated L. johnsonii XS4 from the gastrointestinal mucosa of
healthy laboratory pigs. In previous studies, they demonstrated a high resistance of
L. johnsonii XS4 to hydrochloric acid and bile salts and an antagonistic effect against
the most common pathogens in culture (S. aureus, E. coli, S. Enterica). The sows were
supplemented with freeze-dried L. johnsonii XS4 from day 90 of gestation to day 25 of
lactation. It was noted that the supplementation resulted in an increase in the number of
piglets weaned from the sow and an increase in litter weight by 14.45% compared to the
control. In the supplemented group, there was a lower loss of backfat during lactation
than in the control group, but it was not statistically significant. The obtained results
indicate a positive effect on the production efficiency of the sows and the obtained litter.
L. johnsonii XS4 has been presented as a promising alternative to the use of antibiotics in
feed [76]. In another study, 295 LAB strains were tested, and three strains (Limosilactobacillus
reuteri P7, Lactobacillus amylovorus P8, and L. johnsonii P15) with high growth-inhibitory
activity for enterotoxigenic E. coli K88 were selected. The strains had a positive effect on
the reproductive performance of sows and the growth of weaned piglets and reduced the
occurrence of diarrhea [77].
In contrast, supplementation with E. faecium DSM 7134 increased food digestibility,
gross weight, and gross energy. Moreover, in this case, the supplementation affected the
sow’s litter. The mortality of weaned pigs decreased, and weight gain of the piglets was
noticeable. In this experiment, a reduced number of E. coli in feces was demonstrated after
the piglets were weaned [78].
The administration of probiotic mixes to the sow, in this case, composed of L. del-
brueckii subsp. bulgaricus, L. rhamnosus, L. acidophilus, L. plantarum, Streptococcus salivarius
subsp. thermophilus, Bifidobacterium bifidum, E. faecium, Candida pintolopesii, and Aspergillus
oryzae also gave satisfactory results. In this study, no diarrhea occurred in either the sows
or the piglets. An increased concentration of acetic, propionic, and butyric acids in the
Animals 2021, 11, 3431 9 of 20

feces was shown in piglets supplemented with the mix. However, in this case, the supple-
mentation of the sows did not affect the litter weight, but there was a visible change in the
gut environment of the piglets [79]. In the case of using only the dietary supplementation
L. plantarum CAM6 in sows, it had a positive effect on body weight and reduced the occur-
rence of diarrhea in the offspring. The nutritional value of milk in sows improved [80]. The
application of a diet with L. plantarum JL01 for weaned piglets resulted in better digestion
and absorption of fats in the cecum and mediated the metabolism of tryptophan [81,82].
The interest of scientists was also aroused by the influence of a probiotic diet on
bacteria present in the air. The effect of administering feed supplemented with E. faecalis
CICC 23,215 in a piglet house over 60 days of airborne bacterial communities in the house
was investigated. Air and feces samples were tested. The enrichment of air and feces
with Lactobacillus species was shown. E. faecalis reduced the abundance of Proteobacteria,
Acinetobacter sp., Escherichia sp., and Shigella sp. [83].
In another study by Wang et al. (2021), the effect of feeding with feed co-fermented
by B. subtilis CW4 and E. faecium CWEF on lactating sows and newborn piglets was
investigated. It was shown that the quality of the sows’ milk was improved, which resulted
in an increased weight gain in the piglets. There was a reduction in the incidence of
constipation in sows and diarrhea in piglets, which was related to the modulation of the
intestinal microflora. The sows showed better immunity and performance compared to the
control [84]. Another experiment used the positive effects of Bacillus coagulans, oregano oil,
and benzoic acid on the health, physiological, and physical condition of the piglets. An
increased weight gain with a reduced feed ration was obtained in the diet supplemented
with benzoic acid and B. coagulans. The addition of a third component to the diet—oregano
oil—resulted in an increased number of bifidobacteria in the caecum and a decreased
E. coli population in the cecum. Both the supplementation of the two-component and
three-component diets positively influenced intestinal integrity, immunity, and physical
condition of piglets in non-antibiotic breeding [85]. Similarly, in the study by Fu (2021), the
use of B. coagulans and yeast hydrolysate in weaned piglets improved the intestinal barrier
function, which resulted in better weight gain of the piglets. These two options can be used
as alternatives to antibiotic growth promoters [86].
Not only LABs can be used as potential probiotics to prevent diarrhea. Swine com-
mensal E. coli strains are bactericidal and compete in the environment with pathogenic
porcine strains of E. coli. A reduced susceptibility of commensal E. coli to the 34 bacteri-
ocin monoproducers was demonstrated compared to pathogenic E. coli. Finally, in vitro
and in vivo, three potential probiotic E. coli were selected that could be candidates for the
prophylaxis of post-weaning diarrhea [87]. The above information is briefly summarized
in Table 2.

Table 2. Overview of the application of beneficial bacteria in pig and piglet farming in an in vivo study.

Strains Form/Way of Administration Effect References

Okara fermented soy milk with Better meat quality and growth
L. delbrueckii subsp. delbrueckii per os 1 performance; increased level of lactobacilli [68]
TUA4408L/probiotic and Lactococcus
L. delebureckii CCTCC M 207040/probiotic [69]
Reduced crypt depth in the jejunum and
C. butyricum ZJU-F1 and B. per os in feed 1
ileum; increased gut integrity [70]
licheniformis/probiotic
Antagonistic activity against C. perfringens
MP34, E. faecalis MP42, S. aureus MP83,
Streptococcus suis MP205, Trueperella
L. salivarius MP100/probiotic per os 1,3 pyogenes MP214, E. coli MP73 (F4) and [71]
MP77 (F18), S. Typhimurium MP55, and
Klebsiella pneumoniae MP87
Lactobacillus gasseri LA39 and
Limosilactobacillus frumenti/probiotic feces and saline solutions/per os 2 Prevent diarrhea after weaning [72]

Increased level of lactobacilli and

1 reduction level of Bacteroides and
L. salivarius 144 and 160/probiotic per os Fibrobacter in the gastrointestinal tract; [73]
reduced diarrhea
Animals 2021, 11, 3431 10 of 20

Table 2. Cont.

Strains Form/Way of Administration Effect References

Antagonistic activity against E. coli,
L. plantarum SC01/probiotic Microcapsulation/per os 2 S. aureus, B. subtilis, Salmonella sp., and [74]
L. monocytogenes
Improvement of intestinal integrity,
elongation of intestinal villi in the jejunum,
L. plantarum 22F and 25F, and P. acidilactici Spray drying the appearance of microorganisms
[75]
72N/probiotic microencapsulation/per os 2 positively influencing the intestinal
microbiome, and improved growth of
individuals in the rearing cycle
Increased number of weaned piglets
L. johnsonii XS4/probiotic Freeze-dried/per os 3 increased litter weight
[76]

Positive effect on the reproductive

Limosilactobacillus reuteri P7, Lactobacillus performance of sows and the growth of
amylovorus P8, and L. johnsonii per os 3 [77]
P15/probiotic weaned piglets and reduced the
occurrence of diarrhea
Reduced level of E. coli in feces after
E. faecium DSM 7134 per os 3 weaning the piglets [78]

L. delbrueckii subsp. bulgaricus,

L. rhamnosus, L. acidophilus, L. plantarum,
Streptococcus salivarius subsp. thermophilus, Increased concentration of acetic,
per os 1,3 propionic, and butyric acids in the feces [79]
Bifidobacterium bifidum, E. faecium, Candida
pintolopesii, and Aspergillus
oryzae/probiotic
L. plantarum CAM6/probiotic per os 3 Improved nutritional value of milk [80]
B. subtilis CW4 and E. faecium Improvement in the quality of sows’ milk;
CWEF/probiotic per os 1,3 better immunity at sows
[84]

Increased carcass yield; improved meat

L. reuteri 1/probiotic freeze-dried/per os 2 quality and flavor
[88]

B. subtilis WB800 per os 1 Enhanced respiratory immunity [89]

The experiment was carried out on 1—piglets, 2—pigs, 3—sows.

4. Prevention of Udder Infections in Cattle with Particular Emphasis on Bacterioci

Like-Substance
Inflammation of a cow’s udder, another name mastitis, is caused by physical trauma,
chemical irritation, or bacterial infection. Mastitis can be identified in two types: clinical and
subclinical. Clinical mastitis manifests itself with local and systemic symptoms. Redness,
swelling, pain in the udder area, decreased appetite, increased temperature, reduced
milk production, and a change in milk compounds are visible [90]. Subclinical mastitis
does not alter the udder. Abnormal changes in milk are visible, such as an increased
population of bacteria, a change in quality and compounds, reduced milk production, and
an increased number of somatic cells [91]. Inflammation of the udder tissue has a negative
impact on economical milk production and the animal’s welfare due to pathologies causing
edema, swelling, pain, inflammation, or udder fibrosis [92] and to a reduced reproductive
efficiency [93]. Disease control is hindered by the causes of multifactorial occurrence
and involving a large number of pathogens [94]. The main pathogens inducing mastitis
are S. aureus, Streptococcus uberis, and Streptococcus dysgalactiae [95–97] and more often
isolated strains of E. coli [98]. The treatment consists of identifying the pathogen and then
administering an appropriate intramammary antibiotic [99]. Due to the excessive use of
antibiotics, bacterial resistance increases and treatment efficacy decreases, so there is a
growing interest in replacement therapies without antibiotics [100].
One experiment reviewed a large bank of engineered nisin modifications and discov-
ered three new variants of nisin A M17Q, nisin A HTK, and nisin A T2L. These variants
showed antibacterial activity against S. aureus strains associated with bovine mastitis. It
has been shown to reduce the growth inhibition of commensal bacteria naturally occurring
in milk, such as lactobacilli and lactococci. This is the next step in the development of
substitutes for antibiotic therapies on farms [101].
Animals 2021, 11, 3431 11 of 20

Godoy-Santos et al. (2019) isolated the bovicin HC5 bacteriocin from the rumen bacte-
ria Streptococcus equinus HC5. Bovicin HC5 is a lantibiotic with the ability to bind to lipid II
in the cytoplasm such as nisin [102]. Other studies showed a bovicin HC5 bactericidal effect
against L. monocytogenes, Salmonella Typhimurium, and some species of Clostridium and
Bacillus [103,104]. The antibacterial effect of bovicin HC5 was tested in isolated strains from
animals with mastitis. There were: S. aureus (99 strains), coagulated negative Staphylococcus
sp. (CNS) (44 strains), Streptococcus agalactiae (71 strains), Streptococcus bovis (22 strains),
S. uberis (20 strains), and E. coli (20 strains). Bovicin HC5 has inhibited the growth of most
Streptococcus and Staphylococcus species. A total of 276 pathogenic isolates were tested.
Some 18% of the isolates were not susceptible to the bacteriocin. None of the E. coli strains
showed sensitivity to bovicin HC5. In the case of S. aureus, the pathogen most often respon-
sible for the occurrence of mastitis in cows, as many as 95% of isolates showed the highest
sensitivity to bovicin HC5 [102].
Due to the high costs of producing pure bacteriocins, other scientists decided to test
the effect of a preparation containing live Lactococcus lactis DPC3147 cultures [105]. The
effectiveness of the product was compared with the commercial antibiotic, Terrexine TM,
which is used to treat cows with clinical and subclinical signs of mastitis. In cows with
clinical symptoms, inflammation and/or malaise, clots, pathogens in milk, and poor milk
production occurred. Cows infected with S. aureus and treated with biopreparation showed
a cure rate of 45%, and cows treated with a commercial antibiotic showed a cure rate
of 50%. The above data show that the biopreparation containing the bacteriocin lacticin
3147 produced by L. lactis DPC3147 shows comparable efficacy to the commercial antibiotic.
Cows treated with the biopreparation showed an increased response of the immune system
and a decrease in the number of somatic cells in milk. Five days after the injection into
the replacement biopreparation, no DPC3147 cells were detected, which indicates that the
animals quickly excrete the “live biopreparation”. The above experience allows us to limit
the administration of antibiotics in the treatment of mastitis in the future and shorten the
withdrawal time of treated animals [105].
On the other hand, one of the in vitro studies demonstrated the bactericidal effect of
the supernatant after the L. lactis ssp. lactis bacterial culture was used against the most
common occurring mastitis pathogen. There were obtained uncleaned bacteriocins from
supernatant after bacterial culture and used on pathogens. In this experiment, Malvisi
et al. (2016) confirmed the presence of the class I bacteriocin, nisin A, in the supernatant
by liquid chromatography. The highest antagonistic activity had L. lactis LL11 and SL153
supernatants after bacterial culture. Malvisi et al. (2016) applied the supernatant containing
nisin A to the bovine mammary epithelial cell line BME-UV1. The stimulation of the
secretion of NAGase and the LZ antibacterial enzymes by the cells was visible without
causing damage or adverse inflammatory reactions, and there was no damage to cell
integrity [106].
One study investigated the antimicrobial activity of 13 specific bacteria isolated in vitro
from a honey bee cultivation against mastitis pathogens. Nine types of Firmicutes were
isolated: Apilactobacillus kunkeei Fhon2, Apilactobacillus apinorum Fhon13, Bombilactobacillus
mellis Hon2, Bombilactobacillus mellifer Bin4, Lactobacillus kullabergensis Biut2, Lactobacillus
kimbladii Hma2, Lactobacillus helsingborgensis Bma5, Lactobacillus melliventris Hma8, and
Lactobacillus apis Hma11 and 4 types of Actinobacteria: Bifidobacterium coryneforme Bma6,
Bifidobacterium asteroides Bin2, Bifidobacterium sp. Bin7, and Bifidobacterium sp. Hma3. Three
of the tested oxalicin-resistant S. uberis were inhibited by the use of a combination of a
honey-based medium and 13 specific bacteria. The same was observed in ampicillin and
trimethoprim-sulfamethoxazole-resistant E. coli isolates. The study demonstrated that
13 specific bacterial symbionts in combination with the heather honey matrix showed an
inhibitory effect on the growth of mastitis pathogens [107].
In one of the experiments carried out by Seon-Gyu Kim et al. (2019), synergistic
inhibition of the growth of S. aureus KCTC 3881 by bacteriocin and bacteriophage was used.
The bacteriocin was isolated from L. lactis CJNU 3001. For comparison, bacteriocin and
Animals 2021, 11, 3431 12 of 20

bacteriophage activity were also tested separately. Pure bacteriocins showed antagonistic
activity against S. aureus depending on the dose. Visible effects were obtained at concentra-
tions of 50 and 100 AU/mL. The treatment of S. aureus with a dose of 1 MOI (multiplicity
of infection) of the SAP84 bacteriophage showed a viable cell count of 5.7 Log CFU/mL.
The combined action of phage (0.1 MOI) and bacteriocin (100 AU/mL) showed a reduction
in the number of viable S. aureus cells to 3.3 Log CFU/mL. The combination of bacteriocin
and bacteriophages may be a promising strategy to combat pathogens not only associated
with mastitis in cattle [108].
One of the metabolites produced by LAB is lactic acid [38,109]. Chotigarpa et al. (2018,
2019) performed the time-killing analysis of rice gel with 5% (v/v) lactic acid on E. coli
ATCC 25,922 and E. coli field strains (teat skin samples from healthy dairy cows after
washing udder) in the time interval of 0 to 60 min. The study showed an inhibitory effect
on the growth of E. coli, which means that created gel can reduce the number of pathogenic
bacteria on the cows’ teats. The gel can be an alternative to the antibiotics used and in the
prevention of mastitis. Additionally, the minimum inhibitory concentration (MIC) and
minimum bactericidal concentration (MBC) of pure lactic acid were checked for E. coli
strains. The MIC and MBC were 0.5% lactic acid [110,111].
The analysis of feces in four dairy cows showed the presence of the following Lacto-
bacillus strains: Lactobacillus gasseri, Limosilactobacillus reuteri, and Ligilactobacillus salivarius.
In vitro, the supernatants of these strain cultures showed bactericidal activity against Es-
cherichia coli O157:H7, Mycobacterium avium ssp. paratuberculosis, and the Salmonella species
(Salmonella enteritidis, Salmonella typhimurium, and Salmonella Dublin). They also showed
a low risk of lateral transfer of antibiotic-resistant genes despite showing resistance to
streptomycin (L. gasseri) and kanamycin (L. salivarius). In an ex vivo study, they showed
adherence to bovine intestinal epithelium cells. The use of such fecal isolates may be a
species-specific probiotic for cattle [112]. The above information is briefly summarized in
Table 3.

Table 3. Overview of the application of potential probiotic bacteria in bovine farming in an in vitro study.

Probiotic/Postbiotic Form/Way of Administration Effect References

Nisin derivatives/postbiotic Solution Antibacterial activity against S. aureus [101]
Bactericidal effect against L. monocytogenes,
Bovicin HC5/postbiotic Solution Salmonella Typhimurium, and some [103,104]
species of Clostridium and Bacillus
Bactericidal effect against the most
L. lactis LL11 and SL153/postbiotic Supernatant [106]
common occurring mastitis pathogen
Bacteriocin from L. lactis CJNU Inhibition of the growth of S. aureus
Bacteriophage [108]
3001/postbiotic KCTC 3881
Lactobacillus gasseri, Limosilactobacillus Bactericidal activity against Escherichia coli
reuteri, and Ligilactobacillus Supernatant O157:H7, Mycobacterium avium ssp. [112]
salivarius/postbiotic paratuberculosis, and Salmonella species
Inhibition of the growth of S. aureus;
L. gasseri LA806 Live and heat-inactived [113]
barrier and immunomodulatory

5. Antibiotics—Future Legal Regulations

In order to counteract the phenomenon of antibiotic resistance and protect broadly
understood public health, comprehensive actions, including legal ones, are necessary. In
reference to the data provided by OECD (Organisation for Economic Cooperation and
Development), it is estimated that about 700,000 deaths may be caused globally each year
by AMR (antimicrobial resistance). Compared to a world with no AMR, the economic
impact associated with current rates of AMR may reach about 0.03% of GDP in 2020 in
OECD countries, 0.07% in 2030, and 0.16% in 2050. This would result in a cumulative
loss of about USD 2.9 trillion (NB: in the quoted report, the amount ‘trillion’ means
1012—‘Council conclusions on the next steps under a One Health approach to combat
Animals 2021, 11, 3431 13 of 20

antimicrobial resistance’ (2016/C 269/05), Official Journal of the European Union 23.7.2016)
by 2050 [114]. Moreover, as shown by data published by the European Medicines Agency,
the use of antibiotics in Europe is more than twice as high in the treatment of animals as in
humans. From 2011 to 2014, the use of antibiotics in agriculture increased by 23% [115].
The basic legal act in the European Union in the field of the use of antibiotics, which will
come into force on 28 January 2022, is Regulation (EU) 2019/6 of The European Parliament
and of the Council on 11 December 2018 on veterinary medicinal products, and repealing
Directive 2001/82/EC [116]. The above normative act will be directly applicable in all
Member States, without the obligation to implement it into the national legal order.
According to the above-mentioned legal act, in Article 4 point 12 of the Regulation
(EU), ‘antibiotic’ means any substance with a direct action on bacteria that is used for
treatment or prevention of infections or infectious diseases. The antibiotic also fits into
the broader definition adopted by the EU legislator, the so-called ‘antimicrobial’, which
means any substance with a direct action on micro-organisms used for the treatment or
prevention of infections or infectious diseases, including antibiotics, antivirals, antifungals
and antiprotozoals) [116]. It is also worth recalling the broadest definition in terms of the
analyzed regulation (Vide Article 4 point 1) [116]: ‘veterinary medicinal product’ means
any substance or combination of substances which fulfils at least one of the following
conditions:
(a) it is presented as having properties for treating or preventing disease in animals;
(b) its purpose is to be used in or administered to animals with a view to restoring,
correcting, or modifying physiological functions by exerting a pharmacological, im-
munological, or metabolic action;
(c) its purpose is to be used in animals with a view to making a medical diagnosis;
(d) its purpose is to be used for euthanasia of animals.
Of key importance for the issue discussed is Art. 107 of the Regulation, which expressis
verbis regulates the issues of the use of antimicrobial medicinal products. It shows, inter
alia, that antimicrobial medicinal products cannot be used:
(a) antimicrobial medicinal products shall not be applied routinely nor used to compen-
sate for poor hygiene, inadequate animal husbandry, or lack of care or to compensate
for poor farm management;
(b) in animals for the purpose of promoting growth nor to increase yield;
(c) prophylaxis other than in exceptional cases for the administration to an individual
animal or a restricted number of animals when the risk of an infection or of an
infectious disease is very high, and the consequences are likely to be severe. In such
cases, the use of antibiotic medicinal products for prophylaxis shall be limited to the
administration to an individual animal only, under the conditions laid down in the
first subparagraph.
A Member State may further restrict or prohibit the use of certain antimicrobials in
animals on its territory if the administration of such antimicrobials to animals is contrary
to the implementation of a national policy on prudent use of antimicrobials.
Antimicrobial medicinal products shall be used for metaphylaxis only when the risk of
spread of an infection or of an infectious disease in the group of animals is high and where
no other appropriate alternatives are available. Member States may provide guidance
regarding such other appropriate alternatives and shall actively support the development
and application of guidelines which promote the understanding of risk factors associated
with metaphylaxis and include criteria for its initiation (Vide Article 107 Section 4) [116].
Antimicrobial medicinal products should not be used for prophylaxis other than in well-
defined cases for the administration to an individual animal or restricted number of animals
when the risk for infection is very high or its consequences are likely to be severe (Vide
Article 107 Section 3) [116]. Member States should be able to allow the exceptional use of
veterinary medicinal products without a marketing authorization where it is necessary
to respond to Union-listed diseases or emerging diseases and where the health situation
Animals 2021, 11, 3431 14 of 20

in a Member State so requires (Vide Section 26) [116]. It is worth emphasizing that the
consequences of animal diseases, and at the same time the measures needed to combat
them, can cause enormous damage to the entire population of animals, their owners, and
thus the economy, and ultimately public health.
In order to fulfil the above-mentioned obligations as well as a whole range of other
legal regulations regulated in the analyzed regulation Art. 107 Section 3, said Member
States shall ensure that adequate financial resources are available to provide the staff and
other resources necessary for the competent authorities to carry out the activities required
by this Regulation [116].
The considerations carried out are only indicative, they undoubtedly prove that the
widely understood processes of enactment, but above all the application of law, have a
huge impact on the protection and guarantee of the highest level of public health and
animal health protection as well as environmental protection. Otherwise, misuse (overuse
of antibiotics) will lead to the spread of resistant pathogens, generally in animals, plants,
and the environment and ultimately also in humans, representing one of the most serious
global threats to public health.
The new legal regulation in the European Union does not limit the treatment of sick
animals by administering them antibiotics, but its ratio legis excludes prophylactic treatment
of the entire herd, with only a few diagnosed sick animals in the group. In other words,
the EU legislator prohibits the massive and preventive use of antibiotics, e.g., in animal
feed, water, etc. only for economic purposes. It should also be remembered that only
effective supervision and control activities as well as monitoring studies of individual
public administration bodies in all EU Member States will allow for a reliable assessment of
the legitimacy and correctness of the use of antibiotics, and thus guarantee the protection
of consumers against the consequences of their improper use.
The observation of the practice proves that in addition to the appropriate legal tools
and organizational solutions, including the development of the principles of rational and
safe use of antibiotics, the appropriate staffing of government and local government admin-
istration bodies to perform the above-mentioned duties is of key importance. Otherwise,
staff shortages, insufficient level of financing, and high workloads will make the above-
mentioned tasks ineffective or perhaps more accurately simulated, due to staff fluctuations.

6. Conclusions
The greatest discovery of the 20th century was the control of many diseases, both
animal and human with antibiotics. Unfortunately, the overuse of antibiotics has resulted
in the appearance of drug-resistant pathogenic bacteria in veterinary practice and medicine.
The main goal of large-scale livestock breeders is to intensify production and reduce
mortality in the herd. An easy way to do this is to use antibiotics. Increased animal
production keeps animals crowded, which facilitates the transmission of various diseases.
As shown in the review, the use of probiotics on farms can naturally bring about a balance
of gut microbes and reduce the growth of pathogens in broiler, pig, piglet, and cattle
slaughter farms. Based on the review of the latest articles from 2021, an increase in interest
in the effects caused by B. subtilis can be noticed (Table 1). The results indicate that it could
be a substitute for antibiotics and an effective growth promoter in broiler breeding [55–58].
The use of probiotics is of increasing importance for the alleviation of weight and immunity
deficiencies as well as for lowering the mortality ratio in broiler farms [117]. The discussed
studies show the positive effect of LAB and its metabolites on the welfare of farm animals.
The limitations of antibiotics in farms have a positive effect on the environment and living
organisms, including humans. The application of the new regulations in the law, which will
come into force in 2022, is to limit completely the use of antibiotics for disease prevention
before they appear in the herd. This is the next step to reduce the occurrence of resistance
effects among pathogenic bacteria.

Author Contributions: D.Z., conceptualization, writing—original draft preparation, review, editing,

and Tables 1–3 preparation; A.N., conceptualization, supervision, Figure 1 preparation, review,
Animals 2021, 11, 3431 15 of 20

and editing; I.N., writing—original draft preparation (paragraph 5: Antibiotics—Future Legal

Regulations); and E.M.-P., conceptualization, supervision, and editing. All authors have read and
agreed to the published version of the manuscript.
Funding: This research was funded by European Regional Development Fund, Poland, grant number
RPLD.01.02.02-10-0005/16-00 and RPLD.01.02.02-10-0029/18-00.
Institutional Review Board Statement: Not applicable.
Data Availability Statement: Not applicable.
Acknowledgments: This work paper was completed while the first author was the Doctoral Candi-
date in the Interdisciplinary Doctoral School at the Lodz University of Technology, Poland.
Conflicts of Interest: The authors declare no conflict of interests.

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