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Antibiotics often supplemented in feed, used as a growth promoter may cause their residual effect in animal produce and also trigger antibiotic resistance in bacteria, which is of serious concern among swine farming entrepreneurs. As an alternative, supplementing probiotics gained interest in recent years. Lactobacillus being most commonly used probiotic agent which improves growth performance, feed conversion efficiency, nutrient utilization, intestinal microbiota, gut health and regulate immune system in pigs. The characteristics of Lactobacillus spp. and their probiotic effects in swine production are reviewed here under.
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The use of Lactobacillus as an alternative of antibiotic growth promoters in pigs: a
review
Runjun Dowarah, A.K. Verma, Neeta Agarwal
PII: S2405-6545(16)30148-2
DOI: 10.1016/j.aninu.2016.11.002
Reference: ANINU 125
To appear in: Animal Nutrition Journal
Received Date: 19 August 2016
Revised Date: 28 October 2016
Accepted Date: 5 November 2016
Please cite this article as: Dowarah R, Verma AK, Agarwal N, The use of Lactobacillus as an alternative
of antibiotic growth promoters in pigs: a review, Animal Nutrition Journal (2016), doi: 10.1016/
j.aninu.2016.11.002.
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The use of Lactobacillus as an alternative of antibiotic growth promoters in pigs: a review
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Runjun Dowarah
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, A.K. Verma and Neeta Agarwal
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CAFT in Animal Nutrition, Indian Veterinary Research Institute, Izatnagar-243122, India
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*Corresponding author.
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E-mail address: runjundowarah03@gmail.com
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Abstract
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Antibiotics often supplemented in feed, used as a growth promoter may cause their
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residual effect in animal produce and also trigger antibiotic resistance in bacteria, which is of
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serious concern among swine farming entrepreneurs. As an alternative, supplementing probiotics
23
gained interest in recent years. Lactobacillus being most commonly used probiotic agent which
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improves growth performance, feed conversion efficiency, nutrient utilization, intestinal
25
microbiota, gut health and regulate immune system in pigs. The characteristics of Lactobacillus
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spp. and their probiotic effects in swine production are reviewed here under.
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Key words: Diarrhea score, Gut microbiota, Lactobacilli, Probiotics
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1 Introduction
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Various stress factors including nutritional, environmental and weaning etc. at different
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stages of life affect the animal productivity and health. Weaning stress in piglets being the major
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cause for economic loss to pig farmers (Wilson et al., 1989). As the weaned piglets have limited
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digestive capacity, which triggers fermentation of undigested protein by opportunistic pathogens
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(mainly Escherichia coli, Salmonella) normally existing in gastrointestinal tract (GIT) which
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leads to production of branch chain fatty acids (BCFA) and ammonia nitrogen (Garcia et al.,
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2014). The BCFA and NH
3
-N are the toxic metabolites to intestinal mucosa, which damage
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intestinal mucosa thereby ultimately results in diarrhea (Fuller, 1989; Jensen, 1998). This may
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usually causes a reduction in villous height and digestive enzyme activity for the first few days
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after weaning (Pluske et al., 1995). The common practice of supplementing antibiotics in
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livestock for improved animal performance was condemned due to its adverse effects on animal
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as well as human, the ultimate consumer of the animal produce. Since then, it has been the
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greatest challenge to farmers to rear healthy piglets devoid of antibiotics supplementation.
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However, these stress factors in livestock sector need to be addressed for profitable livestock
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farming.
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In this scenario, latest reports indicate probiotic supplementation in swine seems to be
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better alternative for antibiotic use addressing the safe animal produce as well as to combat
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economic losses in pig farming. The term “Probiotics” is derived from a Greek word ‘biotikos’
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meaning ‘for live’, which was first coined by Parker (1974) and define as the live
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microorganisms, when they were administered in adequate amounts, confer a health benefits on
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the host (FAO/WHO, 2002). At present, probiotics are classified by the US Food and Drug
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Administration as generally recognized as safe (GRAS) ingredients. Among various probiotic
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bacteria, Lactobacillus is most commonly used probiotic agent (McCony and Gilliland, 2007).
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Lactobacilli are gram-positive, non-motile, non-spore forming, acid-tolerant, non-respiring rod
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shaped (bacillus), or spherical (coccus) bacteria which produce lactic acids as the major
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metabolic end-product of carbohydrate fermentation (Cho et al., 2009). In farm animal they
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confer good intestinal health by stimulating the growth of a healthy microbiota (Walter, 2008),
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preventing intestinal colonization of enteric pathogens (Huang et al., 2004; Lee et al., 2012),
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reduced faecal noxious gas emission (Hong et al., 2002), production of antimicrobial substances,
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antibiotic resistance patterns, improving digestive ability and antibody mediated immune
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response, and demonstrable efficacy and safety (Wang et al., 2012; Hou et al., 2015). Probiotics
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are generally host-species specific (Dunne et al., 1999) and believed to be more effective in its
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natural habitat i.e., target species (Kailasapathy and Chin, 2000). However, selection of probiotic
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microbes is one of the most important criteria to get a positive response. The objective of this
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paper is to enlighten the efficacy of various Lactobacillus spp. as probiotic in swine production.
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2 Microorganism commonly used as probiotics
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The microorganisms commonly used as probiotics in livestock are presented in Table 1.
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The genus Lactobacillus has been reported as one of the major bacterial groups found in GIT
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(Dibner and Richards, 2005). Till now, no report was found on safety concerns related to
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Lactobacilli in animals. The genera Bifidobacteria is found to be inhabitant of GIT of both
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animals and humans, which helps in maintaining microbial balance in the GIT by reducing the
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harmful pathogenic microbes thereby, associated with good health status of the host (Huang et al.,
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2004). The genus Enterococcus belongs to the lactic acid bacteria (LAB) group and found
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naturally in food products, which are normal commensals of human and animal. E. faecium is the
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most common in the animal GIT, while in human E. faecium and E. faecalis are prevalent
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(Fisher and Phillip, 2009). These three species of Enterococcus are commonly used probiotics in
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animal/ livestock feeding.
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Bacillus are Gram-positive, spores-forming microorganisms, commonly associated with
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soil, water and air, and present in the intestinal tract due to involuntary ingestion of contaminated
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feed. Though some of the Bacillus species are used as a probiotic, speculation exists for their
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ability to produce toxins (Gaggia et al., 2010).The yeasts are also comprised as a residual
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microbial system of the intestinal microbiome where Saccharomyces cerevisiae is widely present
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in the nature and used in food and beverage industry for its fermentation properties. It is also
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used as a probiotic especially in ruminants and pig feeding (Kumar et al., 2012).
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3 Mode of action of Lactobacilli as probiotic
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Lactobacilli stimulate rapid growth of beneficial microbiota in the GIT which become
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abundant and induce competitive exclusion of pathogenic bacteria either by occupying binding
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sites on intestinal mucosa or competing for nutrients and absorption sites with pathogenic
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bacteria (Malago and Koninkx, 2011; Zhao and Kim, 2015); by rapid utilization of energy source
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which may reduce the log phase of bacterial growth. Most of the enteric pathogens adhere to the
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intestinal epithelium through colonization thereby develop diseases (Walker, 2000).
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Consequently, the probiotic lactobacilli have the ability to adhere the gut epithelium and thus
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compete with pathogens for adhesion receptors i.e., glycol-conjugates (Umesaki et al., 1997).
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Probiotic bacteria produce organic acids, hydrogen peroxide, lactoferrin and bacteriocin which
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may exhibit either bactericidal or bacteriostatic properties (Jin et al., 1997; Pringsulaka et al.,
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2015). Lactobacillus have proven to be capable of acting as immune-modulators by enhancing
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macrophage activity (Perdigon et al., 1986), increasing the local antibody levels (Yasui et al.,
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1989), inducing the production of interferon (De Simone et al., 1986) and activating killer cells
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(Kato et al., 1984). They prevent the proliferation of coliform bacteria thus amine production
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diminishes which produced due to decarboxylation of amino acids by coliform bacteria.
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4 Selection of lactobacilli for feeding as Probiotics
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The followings are the criteria that can be used for the selection of microbial strains as
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probiotics:
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1) Resistance to in vitro/in vivo conditions: they should be resistant to acidic pH and bile
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salt. After administration, the microbes should not be killed by the defense mechanisms of
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the host and should be resistant to the specific conditions occurring in the body.
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2) Origin of the strain: Probiotics are generally host-species specific (Dunne et al., 1999). It
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is believed that probiotic organism is more effective if it is naturally occurring in the target
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species (Kailasapathy and Chin, 2000). The strains should be properly isolated and identified
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before use.
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3) Biosafety: Lactobacillus, Bifidobacteria and Enterococcus are the microbes which fall in
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the category of generally recognized as safe (GRAS) and are most widely used
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microorganisms as probiotics.
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4) Viability/survivability and resistance during processing (e.g., heat tolerance or storage):
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Thermophilic/thermo-tolerant organisms have an advantage as they withstand higher
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temperature during processing and storage.
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However, other criteria might also be considered for selection of mono or multi strains
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bacteria as probiotics like as probiotic-symbiotic interaction, stimulation of healthy microbiota
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and suppression of harmful bacteria. Adopting these predetermined criteria, it could be possible
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to select the best strains of probiotics which could be effective therapeutically and nutritionally.
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5 Mode of feeding probiotics
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Mode of feeding probiotic affects the response of animal to the probiotic feeding.
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Generally cultures are fed either in form of lyophilized powder or live cells. When a lyophilized
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culture fed to the animal, it has to rejuvenate before expressing its biological activity which takes
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some time and also the viability of the culture is not assured. Whereas, when live culture is fed,
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the probiotic starts its biological activity immediately and the viability of the culture is assured.
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Further, cells can lose their viability and metabolic activity during lyophilisation. Therefore, a
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probiotic product is effective only when it contains viable as well as metabolic active cells.
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6 Use of probiotic Lactobacilli spp. for swine production
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Lactobacillus spp. has been reported as one of the major bacterial groups found in
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porcine gastrointestinal tract (Dibner and Richards, 2005). But all the species and strains of
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Lactobacillus cannot be equally effective as probiotic. Different species and even strains of
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species of microbes behave differently (Newbold et al., 1995). Therefore, source of microbe to
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be used as probiotic is one of the important factors to yield good response. A microbe fed
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exogenously as probiotic when enters the GIT has to compete with existing microbiome
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ecosystem in GIT (Verdenelli et al., 2009). These unfavorable circumstances in the GIT may
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hamper the proliferation of the microbe used as probiotic. To combat this problem, it is
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suggested that the microbe isolated from the host animal when used as probiotic gives better
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response. Host specificity was observed among Lactobacilli isolated from human and animal
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sources (Morelli, 2000). Hence, evaluating new sources of probiotics from swine origin is utmost
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necessary to improve pig farming. Lactobacillus acidophilus 30SC isolated from swine showed
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excellent acid resistance and bile tolerance compared to commercial probiotics strains that are
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presently used in dairy products (Oh et al., 2011). The scenario of using LAB as probiotic for
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swine production is illustrated considering recent reports across the world (Figure 1 & Table 2).
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6.1 Improved performance
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In swine, the use of probiotics improves intestinal well-being which leads to improve
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performance. Higher growth rate and improved feed efficiency ratio results in improve
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profitability due to greater output and reduction in overhead costs (Campbell, 1997). However,
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age and weight at weaning are closely related to post-weaning growth rate. The administration of
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probiotics, soon after birth could be effective as probiotic bacteria by enhancing intestinal barrier
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function which restrict colonization of pathogenic bacteria to intestinal mucosa. This is evident
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with better absorption of nutrients and immunoglobulins of the colostrum, enabling better
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sustainability of the piglet, and minor loss of piglets at its first days of life (Abrahao et al., 2004).
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Supplementation of lactobacilli has resulted in improved growth and feed efficiency in nursery
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(Fialho et al., 1998) and weaning (Mishra et al., 2014) piglets.
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A healthy intestinal tract has a dominance of LAB, however, this equilibrium within the
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intestinal tract is troubled when the animal is in stressful condition like castration, weaning, high
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temperature and humidity and change of feed (Jin et al., 1997). This could be improved by
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continuous feeding of lactobacilli, which encourages rapid growth of other beneficial bacteria
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and reduce the growth of pathogenic bacteria by competitive exclusion (Pan and Yu, 2014).
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Feeding of probiotics (Lactobacillus spp.) to weaning piglets resulted in an increased growth rate
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due to high feed intake and better feed conversion ratio. Supplementation of complex probiotics,
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including yeast (Saccharomyces cerevisiae) and Lactobacillus spp. have been reported to
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improve growth performance of weaned piglets (Hong et al., 2002; Chen et al., 2005). Positive
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effects on growth, feed conversion efficiency and nutrient digestibility were also observed in
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grower-finisher pigs with supplementation of probiotic Bacillus subtilis, Saccharomyces
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boulardi and L. acidophilus C3 (Giang et al., 2011). Where, the effect of probiotic (Lactobacillus
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reuteri) in weaning piglets was comparable with antimicrobial growth promoter (Apramycin) in
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gain, feeding efficiency and nutrient digestibility (Ahmed et al., 2014). This might be due to
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increase activity of digestive enzymes like β-galactosidase by the supplementation of lactobacilli,
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which stimulate gastrointestinal peristalsis and promote apparent nutrient digestibility (Wang et
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al., 2011; Zhao and Kim, 2015).
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Effect of probiotic was also observed in sow with increased feed intake, number and
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growth of piglets at weaning (Pollmann et al., 2005) and also improve composition of milk
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(Alexopoulos et al., 2004). There is a lack of study conducted on sow to see the effect of
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lactobacilli as probiotics, so it important to give emphasis on research in this aspect for better
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swine production. Supplementation of L. johnsonii XS4 (previously isolated from
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gastrointestinal tract of sow) from 90
th
day of pregnancy up to weaning day (25th day of
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lactation) significantly increased litter weight at birth, litter weight after 20 days of farrowing,
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the number of piglets at weaning and weaning weight of piglets (Wang et al., 2014).
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6.2 Control of Post-weaning Diarrhea
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At the time of weaning, entero-pathogens takes upper hand as weaning of the piglets
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reduce digestibility of high protein diet which results increase production of BCFA and ammonia
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nitrogen (NH
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-N) leading to more incidence of diarrhea (Garcia et al., 2014). The BCFA and
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NH
3
-N are the main toxic metabolites for intestinal mucosa and tagger of post weaning diarrhea
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in piglets (Williums et al., 2005). E. coli are the major enteropathogen of post-weaning diarrhea
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and causes 26% cases of neonatal diarrhea (Hoefling, 1989). Addition of lactobacilli as probiotic
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in the diet results beneficial fermentation resulting increased concentration of short chin fatty
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acids and lactic acid in GIT. These may reduce pH of the gut which in turn decrease growth of
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opportunistic entero-pathogens as they need alkaline medium to proper growth and
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multiplications.
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6.3 Effect in intestinal microflora and gut health
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Microflora in the digestive system plays a very important role in the defense mechanism
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of the body. The major intestinal microflora of pig are Lactobacilli, Bifidobacteria, Streptococci,
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Bacteriodes, Clostridiums perfringes and E. coli may vary with age. One of the important ability
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of stable microflora in gastrointestinal tract is colonization resistance. About 4 to 6 weeks is
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needed to establish a stable microflora in the GIT (Mul and Perry, 1994). However,
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supplementation of Lactobacilli in neonatal piglets helps in early development of stable gut
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microbflora, stimulation of immune system and prevents diarrhea. When piglets are weaned, the
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intestinal microflora of piglets is altered due to dietary and environmental change after weaning
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of piglets (Jensen, 1998). The entero-pathogenic E. coli are markedly increased in the anterior
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small intestine resulting post weaning diarrhea. Oral administration of L. fermentum I5007 in
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formula-fed piglets improved intestinal health and reduced the number of potential entero-
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pathogens like E. coli and Clostridia in neonatal piglets (Liu et al., 2014). Addition of complex
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lactobacilli previously isolated from GIT of piglet (L. gasseri, L. reuteri, L. acidophilus, L.
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fermentum, L. johnsonii and L. mucosae) increased number of lactobacilli and bifidobacterium,
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also reduced E. coli and aerobic bacteria counts in jejunum, ileum, cecum and colon mucosa
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(Huang et al., 2004; Chiang et al., 2015). The effect was also persisted in grower-finisher pigs
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(Giang et al., 2011). Since, lactobacilli produced lactic acid, hydrogen peroxide and lactoferrin
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which may exhibit antagonistic activity against E. coli and Enterobacteria (Li et al., 2015). In
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addition, earlier report also revealed that dietary supplementation of complex of lactic acid
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bacteria increased concentrations of lactic acid and acetic acid in ileum and colon (Giang et al.,
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2010) of weaning piglets, whereas, faecal NH
3
-N and butyric acid concentration was decreased
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in grower-finisher pigs (Chen et al., 2006). The increased concentration of lactic acid in
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supplemented group lowered the gut pH in treated groups as lactic acid act as an acidifying agent
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(Williams et al., 2005; Thu et al., 2011). This indicates that supplementation of various
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Lactobacilli sp. modulates gut microbial profile and thereby affected microbial metabolite
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production which results better gut health.
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6.4 Improved immune status
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The probiotics have the capacity to modulate the immune system of animal by enhancing
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the systematic antibody response to soluble antigens in the serum (Christensen et al., 2002). The
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immune-modulatory effects can even be achieved by dead probiotic bacteria or just probiotics
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derived components like peptidoglycan fragments or DNA. Dietary supplementation with
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probiotics enhanced humoral and cell mediated immune responses (Shu and Gill, 2002) with
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increased the serum concentration of IgM (Dong et al., 2013) and IgG (Ahmed et al., 2014)
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growing pigs. Probiotic supplementation in sow also increased IgG level in colostrum and
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plasma of piglets (Jang et al., 2013). Since, probiotics facilitate the suppression of lymphocyte
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proliferation and cytokine production by T cells which down regulating the expression of pro-
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inflammatory cytokines such as tumor necrosis factor-α (Isolauri et al., 2001). L. fermentum
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enhanced T-cell differentiation, induced cytokine expression in the ileum of E. coli challenged
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piglets with increased pro-inflammatory cytokines and percentage of CD4
+
lymphocyte subset in
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blood (Wang et al., 2009). However, it is difficult to confirm that probiotics contribute
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significantly to the immune system of the host. The main reason behind this caveat is that
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probiotics differ from antibiotics in that they are not intended to eradicate invasive pathogens in
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the gastrointestinal tract. Therefore, such observed improvements or positive effects are always
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hindered due to the animal’s immune status and the various applied situations.
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6.5 Antioxidant status
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An abnormality in the antioxidant defense system can increase the susceptibility of pigs
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to stress, resulting in decreased performance and reduced immune function. As a result of
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incomplete reduction of oxygen, the reactive oxygen are formed which includes superoxide
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anion, hydroxyl radical, hydrogen peroxide and singlet oxygen. A physiological concentration of
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reactive oxygen species (ROS) is required for normal cell function, energy production,
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phagocytosis and intercellular signaling regulation. Early weaning of piglets cause oxidative
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stress by producing excessive quantities of ROS which not only damage proteins, lipids and
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DNA but also decline intestinal antioxidant enzyme activities under NF-κB, p65 and Nrf2/
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Keap1 signals (Yin et al., 2014; Yin et al., 2015). The probiotic cell have defenses mechanisms
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against the damaging effects of ROS by involving both in enzymatic (superoxide dismutase and
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catalase) and non-enzymatic components. The lactic acid bacteria diminish the activity of ROS
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through the production of superoxide dismutase (Stecchini et al., 2001) that converts superoxide
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radicals to oxygen and hydrogen peroxide. Some of the species of LAB may also produce
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catalase, which can destroy hydrogen peroxide at a very high rate that blocks formation of
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peroxyl radicals (Knauf et al., 1992) while some lactobacilli produced non-enzymatic
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antioxidants such as glutathione and thioredoxin to reduce reactive oxygen intermediates (de
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Vos, 1996). Supplementation of Lactobacilli sp. increased serum concentration of superoxide
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dismutase, glutathione peroxidase and catalase in suckling (Cai et al., 2014) and weaning piglets
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(Wang et al., 2012) whereas, total antioxidant capacity, hepatic catalase, muscle superoxide
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dismutase improved in grower-finisher pigs (Wang et al., 2009).
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6.6 Effect on intestinal morphology
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The gastrointestinal tract is the main digestive and absorptive organ in animal. The GIT
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permits the uptake of dietary substances into systemic circulation and it also excludes pathogenic
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compounds simultaneously (Gaskins, 1997). There is a reduction in villous height (villous
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atrophy) and crypt depth at weaning (Pluske et al., 1996). As weaning leads to temporary
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starvation which resulted villous atrophy, reduces mucosal protein content and digestive
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enzymes activity
(
McCracken and Kelly, 1984). Hence, improve feed intake immediately after
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weaning reduced the histological changes of small intestinal morphology (Pluske et al., 1996).
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As feeding of probiotic increased daily feed intake thus it had positive effect on development of
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intestinal epithelium. However, the effects of probiotic on villus height may change depending
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upon species of microorganism. Longer villi (V) height, deeper crypt (C) depth and smaller V: C
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ratio was observed in pigs supplementation with L. acidophilus (Rodrigues et al., 2007), L.
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plantarum (Suo et al., 2012) and P. acidolactici (Giancamillo et al., 2008). However, no change
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in the crypt depth was observed with Enterococcus faecium (Scharek et al., 2007).
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6 Conclusion
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In conclusion, the data available from various studies and application of Lactobacilli in
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swine husbandry clearly indicate that various Lactobacilli spp. have a great potential as an
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alternative of antibiotics. More attention should be paid for utilizing effects of different probiotic
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preparations and corresponding feeding strategy in pigs.
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Conflict of interest
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The authors declare that they have no completing of interests.
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Table 1: List of microorganisms commonly used as probiotics in animal feed
Genus Species
Lactobacillus L. acidophilus
L. casei
L. delbrueckii sub sp. bulgaricus
L. brevis
L. cellobiosus
L. curvatus
L. fermentum
L. plantarum
L. reuteri
L. salivarius sub sp. thermophilus
L. gasseri
Lactococus L. cremoris
L. lactis
Pediococcus P. acidilactici
P. pentosaceus subsp. pentosaceous
Bifidobacterium B. bifidum
B. adolescentis
B. animalis
B. infantis
B. longum
B. pseudolongum
B. thermophilum
Enterococcus E. faecium
Bacillus subtilis
coagulans
cereus
licheniformis
Yeast Saccharomyces cerevisiae
Aspergillus oryzae
Source: Dunne et al., 2001; Sekhon and Jairath, 2010
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Table 2: Effect of feeding different Lactobacillus sp. in various categories of pig
Used group Strain Effect of feeding Reference
Suckling and
Nursery piglets B. pseudolongum & L. acidophilus Improved weight gain, feed conversion and
fecal condition Abe et al., 1995
L. reuteri & Bifidobacterium
pseudolongum Better feed conversion ratio with no
differences in final weight, weight gain and
feed intake
Afonso et al., 2013
L. fermentum Increased average daily gain; reduced
diarrhea incidence and numbers
of Clostridium sp; reduced mRNA
expression of IL-1β in the ileum
Liu et al., 2014
Weaned piglets L. gasseri, L. reuteri, L. acidophilus
& L. fermentum Better growth performance, increased
lactobacilli with decreased E. coli counts
and reduced post weaning diarrhea
Huang et al., 2004
L. reuteri, B. subtillis & B.
licheniformis Increased nutrient digestibility, reduced
faecal Salmonella and E. coli number,
improved serum IgG level
Ahmed et al., 2014
L. gasseri, L. reuteri, L. acidophilus
and L. fermentum Better ADG, ADFI and CP digestibility;
improved serum specific anti-OVA IgG Yu et al., 2008
Grower-Finisher
pigs
L. plantarum
L. fermentum Enhanced superoxide dismutase,
glutathione peroxidase and catalase Wang et al., 2012
Wang et al., 2009
L. reuteri, L. salivarius, L.
plantarum and Yeast complex Improved ADG, ADFI and gain/feed,
digestibility of DM and N Shon et al., 2005
E. faecium, L. acidophilus,
Pediococcus pentosaceus and L.
fermentum
Improved growth, FCR, digestibility of
nutrients Giang et al., 2011
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... Additionally, some probiotics used in animal production have increased significantly over 10 years. Particularly, both Bacillus and lactic acid bacteria (LAB) are considered effective and safe alternatives to antibiotics for animal production due to their high stability in vivo (Dowarah et al., 2017;Mingmongkolchai and Panbangred, 2018). Bacillus belongs to Gram-positive bacteria and can form spores, which are favorable for long-term storage; Bacillus can improve growth performance, immunity function, and gut health in animals (Grant et al., 2018;Ramlucken et al., 2020). ...
... Meanwhile, CFS of LAB was more efficient than CCE, which can contribute to declining the burden of infection caused by ARB in farms (Musliu et al., 2021). The crude extract isolation of herbal plants and the cultivation of probiotics is a simple process that does not require the use of any sophisticated equipment and techniques, thus they may be recommended as ones of the most promising alternatives to antibiotics and used in the treatment of MDR pathogens in both local and modern animal farms to reduce antibiotic usage (Dowarah et al., 2017;Huang et al., 2018;Abdallah et al., 2019). ...
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The overuse of antibiotics in food animals has led to the development of bacterial resistance and the widespread of resistant bacteria in the world. Antibiotic-resistant bacteria (ARB) and antibiotic-resistant genes (ARGs) in food animals are currently considered emerging contaminants, which are a serious threat to public health globally. The current situation of ARB and ARGs from food animal farms, manure, and the wastewater was firstly covered in this review. Potential risks to public health were also highlighted, as well as strategies (including novel technologies, alternatives, and administration) to fight against bacterial resistance. This review can provide an avenue for further research, development, and application of novel antibacterial agents to reduce the adverse effects of antibiotic resistance in food animal farms.
... The most common probiotics are lactic acid-producing bacteria belonging to the genera Lactobacillus, Bifidobacterium, Enterococcus and yeasts such as Saccharomyces sp. (Dowarah et al., 2017a;Kiros et al., 2018). Hypothesised mechanisms of action of probiotics are reduction of the pH (unfriendly environment for intestinal pathogens), the adhesion to the intestinal epithelial surface to prevent pathogen attachment. ...
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... In lactating animals, milk production, weight gain, and nutrient digestibility depend on microbial population present in their gastrointestinal tract (GIT) (Jami and Mizrahi, 2012). It is recommended to improve microbial balance in GIT by the application of microbial growth promoters (probiotics) (Dowarah et al., 2017). ...
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... ② Adhesion to the GIT wall to prevent colonization by pathogenic microorganisms: The majority of enteric pathogens might colonize the intestinal epithelium and cause disease as a result [117]. As a result, Lactobacillus can adhere to the gut epithelium and compete with pathogens for adhesion receptors, such as glycoconjugates [118]. The Lactobacillus and Bifidobacterium have hydrophobic surface layer proteins that assist the bacteria non-specifically by adhering to the animal cell surface [119]. ...
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A study evaluated the effects of adding multi-enzyme mixture to diets deficient in net energy (NE), standardized ileal digestible (SID) amino acids (AA), standardized total tract digestible (STTD) P, and Ca on growth performance, bone mineralization, nutrient digestibility and fecal microbial composition of grow-finish pigs. A total of 300 pigs (initial body weight [BW] = 29.2 kg) were housed by sex and BW in 45 pens of 7 or 6 pigs and fed 5 diets in a randomized complete block design. Diets were positive control (PC); and negative control 1 (NC1) or negative control 2 (NC2) without or with multi-enzyme mixture. The multi-enzyme mixture supplied at least 1,800, 1,244, 6,600, and 1,000 units of xylanase, β -glucanase, arabinofuranosidase and phytase per kilogram of diet, respectively. The PC was adequate in all nutrients. The NC1 diet had lower content NE, SID AA, STTD P and Ca than PC diet by about 7%, 7%, and 32%, and 13%, respectively. The NC2 diet had lower NE, SID AA, STTD P and Ca than PC diet by 7%, 7%, and 50%, and 22%, respectively. The diets were fed in 4 phases based on BW; Phase 1: 29-45 kg, Phase 2: 45-70 kg, Phase 3: 70-90 kg, and Phase 4: 90-120 kg. Nutrient digestibility, bone mineralization, and fecal microbial composition were determined at the end of Phase 1. Pigs fed PC diet had greater (P < 0.05) overall G:F than those fed NC1 diet or NC2 diet. Multi-enzyme mixture increased (P < 0.05) overall G:F, but the G:F of the multi-enzyme mixture-supplemented diets did not reach (P < 0.05) that of PC diet. Multi-enzyme mixture tended to increase (P = 0.08) femur breaking strength. Multi-enzyme mixture increased (P < 0.05) the ATTD of GE for the NC2 diet, but unaffected the ATTD of GE for the NC1 diet. Multie-nzyme mixture decreased (P < 0.05) the relative abundance of the Cyanobacteria and increased (P < 0.05) relative abundance of Butyricicoccus in feces. Thus, the NE, SID AA, STTD P, and Ca could be lowered by about 7%, 7%, 49%, and 22%, respectively, in multi-enzyme mixture-supplemented diets without negative effects on bone mineralization of grow-finish pigs. However, multi-enzyme mixture supplementation may not fully restore G:F of the grow-finish pigs fed diets that have lower NE and SID AA contents than recommended by 7%. Since an increase in content of Butyricicoccus in intestine is associated with improved gut health, addition of the multi-enzyme mixture in diets for pigs can additionally improve their gut health.
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In order to determine the effects of dietary supplementation of two different probiotics (Saccharomyces cerevisiae and Lactobacillus acidophilus) on growth performance, nutrient digestibility and faecal microbial count in weaned piglets (28d of age), 36 crossbred (Landrace x Local) piglets were allocated into three treatments on the basis of the BW in a completely randomized design. Each treatment was comprised of four replicates with three piglets in each. Dietary treatments were basal diet without any probiotic (Control), or with 10% of the basal diet replaced by feed fermented with either S. cerevisiae NCDC-49 (SC; 3-5x10(6) cfu/g) or L. acidophilus NCDC-15 (LA; 2-3x10(9) cfu/g). Supplementation of SC or LA to piglets improved (P<0.05) BW gain and ADG. Digestibility of nutrients, however remained similar (P>0.05) among the groups. However, piglets in SC and LA groups showed better (P<0.05) FCR compared to Control. Yeast count and lactobacillus counts were higher (P<0.01) in both SC and LA groups as compared to Control accompanying a concomitant reduction (P<0.01) in coliform count. The results indicated that dietary supplementation of S. cervisiae or L. acidophilus has a positive effect on the performance of weaned piglets.
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Two probiotic strains, Lactobacillus johnsonii x-1d-2 and Lactobacillus mucosae x-4w-1, originally isolated from piglet feces, have been demonstrated to possess antimicrobial activities, antibiotic resistances and interleukin-6 induction ability in RAW 267.4 macrophages in our previous study. These characteristics make L. johnsonii x-1d-2 and L. mucosae x-4w-1 good candidates for application in feed probiotics. In this study, soybeal meal, molasses and sodium acetate were selected to optimize the growth medium for cultivation of L. johnsonii x-1d-2 and L. mucosae x-4w-1. These two strains were then freeze-dried and mixed into the basal diet to feed the weaned piglets. The effects of L. johnsonii x-1d-2 and L. mucosae x-4w-1 on the growth performance and fecal microflora of weaned piglets were investigated. The results showed that the bacterial numbers of L. johnsonii x-1d-2 and L. mucosae x-4w-1 reached a maximum of 8.90 and 9.30 log CFU/mL, respectively, when growing in optimal medium consisting of 5.5% (wt/vol) soybean meal, 1.0% (wt/vol) molasses and 1.0% (wt/vol) sodium acetate. The medium cost was 96% lower than the commercial de Man, Rogosa and Sharpe medium. In a further feeding study, the weaned piglets fed basal diet supplemented with freeze-dried probiotic cultures exhibited higher (p<0.05) body weight gain, feed intake, and gain/feed ratio than weaned piglets fed basal diet. Probiotic feeding also increased the numbers of lactobacilli and decreased the numbers of E. coli in the feces of weaned piglets. This study demonstrates that L. johnsonii x-1d-2 and L. mucosae x-4w-1 have high potential to be used as feed additives in the pig industry.
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Probiotics are living microorganisms that provide a wide variety of health benefits to the host when ingested in adequate amounts. The bacterial strains most frequently used as probiotic agents are lactic acid bacteria, such as Lactobacillus reuteri, which is one of the few endogenous Lactobacillus species found in the gastrointestinal tract of vertebrates, including humans, rats, pigs and chickens. L. reuteri is one of the most well documented probiotic species and has been widely utilized as a probiotic in humans and animals for many years. Initially, L. reuteri was used in humans to reduce the incidence and the severity of diarrhea, prevent colic and necrotic enterocolitis, and maintain a functional mucosal barrier. As interest in alternatives to in-feed antibiotics has grown in recent years, some evidence has emerged that probiotics may promote growth, improve the efficiency of feed utilization, prevent diarrhea, and regulate the immune system in pigs. In this review, the characteristics of L. reuteri are described, in order to update the evidence on the efficacy of using L. reuteri in pigs.
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A total of 59 lactic acid bacteria (LAB) strains were isolated from corn stover silage. According to phenotypic and chemotaxonomic characteristics, 16S ribosomal DNA (rDNA) sequences and recA gene polymerase chain reaction amplification, these LAB isolates were identified as five species: Lactobacillus (L.) plantarum subsp. plantarum, Pediococcus pentosaceus, Enterococcus mundtii, Weissella cibaria and Leuconostoc pseudomesenteroides, respectively. Those strains were also screened for antimicrobial activity using a dual-culture agar plate assay. Based on excluding the effects of organic acids and hydrogen peroxide, two L. plantarum subsp. plantarum strains ZZU 203 and 204, which strongly inhibited Salmonella enterica ATCC 43971(T), Micrococcus luteus ATCC 4698(T) and Escherichia coli ATCC 11775(T) were selected for further research on sensitivity of the antimicrobial substance to heat, pH and protease. Cell-free culture supernatants of the two strains exhibited strong heat stability (60 min at 100°C), but the antimicrobial activity was eliminated after treatment at 121°C for 15 min. The antimicrobial substance remained active under acidic condition (pH 2.0 to 6.0), but became inactive under neutral and alkaline condition (pH 7.0 to 9.0). In addition, the antimicrobial activities of these two strains decreased remarkably after digestion by protease K. These results preliminarily suggest that the desirable antimicrobial activity of strains ZZU 203 and 204 is the result of the production of a bacteriocin-like substance, and these two strains with antimicrobial activity could be used as silage additives to inhibit proliferation of unwanted microorganism during ensiling and preserve nutrients of silage. The nature of the antimicrobial substances is being investigated in our laboratory.
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The diverse collection of microorganisms colonising the healthy gastrointestinal tract of pigs, referred to as the microbiota, plays an essential role not only for the well-being of the animal, but also for animal nutrition and performance and for the quality of animal products. A number of naturally-occurring and artificial factors has been shown to affect the composition and activity of the microbiota in the gastrointestinal tract of pigs, these include: diet composition, growth promoting antibiotics, copper, use of probiotics, specific carbohydrates, organic acids and fermented feed. It is generally accepted that the microbiota in the small intestine competes with the host animal for easily digestible nutrients and at the same time produces toxic compounds. A pronounced microbial fermentation occurs in the stomach and small intestine in young piglets. Results have shown that approximately equal amounts of organic acids were produced in the three compartments: stomach, small intestine and large intestine. Further, experiments have shown that as much as 6% of the net energy in the pig diet could be lost due to microbial fermentation in the stomach and small intestine. On the other hand it has been shown that on a normal Danish pig diet, 16.4% of the total energy supply for the pig is achived from microbial fermentation in the large intestine. However, the microbiota in the gastrointestinal tract is unstable the first week after weaning and it takes 2 to 3 weeks after weaning before the fermentation capacity of the microbiota in the hindgut has developed. Use of growth promoting antibiotics in the feed is widespread in pig production. However, the use of the antibiotics avoparcin and virginiamycin as growth promoters in animal feed has been associated with an increase in resistance of bacteria to therapeutic agents and a fear that this could reduce the ability to treat diseased humans. This has caused an increased awareness of the use of antibiotics and a general wish to reduce the use of antibiotic growth promoters. Probiotics, organic acids and specific carbohydrates (yeast cell walls) are often suggested as alternatives to the use of antibiotic growth promoters. However, due to their relative high prices and the variability and unpredictability of their effect, the use of these products is financially questionable in practical pig production. One way to solve this problem may be the use of fermented liquid feed. Fermented liquid feed is characterised by a high number of lactic acid bacteria, high number of yeast and high concentration of lactic acid, and several investigations has shown fermented liquid feed to improve growth performance in pigs and to established a prophylactic barrier against gastrointestinal disorders. In general there is no doubt that the effect of feed additives are greatest in young animals where they have been found to improve growth performance and to reduce scouring and neonatal mortality. The present paper review our current state of knowledge about how various feed additives affect the gastrointestinal ecosystem.
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The study determined the effects of Lactobacillus fermentum I5007 administered early in the life of piglets on their growth, faecal microflora, immune index and antioxidant activity. Twelve litters of 3-day-old crossbreed piglets were randomly assigned to either of two treatments: piglets in the control group were orally administered 2 ml saline per day per piglet from day 3 to day 5 of life; piglets in the probiotics group were given 2 ml rejuvenation liquid of L. fermentum spray-dried powder (≥ 109 CFU ml-1). Compared with the control group, oral administration of L. fermentum tended to increase the weight gain (P < 0.10) of piglets from day 3 until day 17 of life, and significantly (P < 0.05) increased the number of Lactobacillus in faeces, the concentrations of IgM, IgG, IgA and glutathione, the activity of glutathione peroxidase, and the total antioxidant capacity in plasma of piglets on day 17. In conclusion, oral administration of L. fermentum to piglets early in life could be beneficial by establishing appropriate intestinal microbial flora.
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