Open access peer-reviewed chapter

The Use of Probiotics in Poultry Production for the Control of Bacterial Infections and Aflatoxins

Written By

Daniel Hernandez-Patlan, Bruno Solis-Cruz, Billy M. Hargis and Guillermo Tellez

Submitted: 04 May 2019 Reviewed: 25 July 2019 Published: 11 September 2019

DOI: 10.5772/intechopen.88817

From the Edited Volume

Prebiotics and Probiotics - Potential Benefits in Nutrition and Health

Edited by Elena Franco-Robles and Joel Ramírez-Emiliano

Chapter metrics overview

1,850 Chapter Downloads

View Full Metrics

Abstract

In intensive poultry production, a large number of antimicrobials are frequently employed to prevent (prophylactic use) and treat (therapeutic use) diseases, as well as for growth promotion (subtherapeutic use), in order to increase productivity. However, it has been reported that the use of antimicrobials at subtherapeutic doses is closely related to the increase in bacterial resistance and with the treatment failure. In addition to antimicrobial resistance, another problem derived from the use of antimicrobials is the presence of residues in animal products. Therefore, these problems and the ban of antimicrobial as growth promoters have prompted the poultry industry to look for alternatives with similar benefits to antibiotics. Among these alternatives, probiotics are one of the most widely studied and interesting groups. Hence, in the present chapter, the effect of probiotics and direct-fed microbial against foodborne pathogens and mycotoxins will be summarized.

Keywords

  • probiotics
  • direct-fed microbial
  • foodborne pathogens
  • antimicrobial resistance
  • aflatoxins

1. Introduction

Since the discovery and application of penicillin in 1940, antibiotics have played an unprecedented role in the prevention, control, and treatment of infectious diseases in both humans and animals [1]. However, in animal production, they have also been used at subtherapeutic doses [2]. It is estimated that the global consumption of antibiotics in animal production could increase by 67% in the coming years [3] mainly because of the growing global demand for animal protein [2, 4]. Although it has been reported that in developed countries the total consumption of antibiotics has decreased by around 4%, consumption of antibiotics in the USA increased slightly [5]. Furthermore, it has been reported that the amount of antibiotics used in animal production in the USA is 100–1000 times higher than human medicine, being used ~80–90% at subtherapeutic doses, and for prophylactic purposes, while the remaining 10–20% at therapeutic doses [6, 7].

The inclusion of antibiotics at subtherapeutic doses into the feed was generalized in the early 1950s, both in the EU and the USA since they could be used to prevent diseases and positively influence the promotion of growth and feed efficiency of animals [3, 8, 9].

Nevertheless, in the last decades, these practices have changed considerably due to the concern of the increase of bacteria resistant to antibiotics, since they can be transmitted zoonotically from animals to humans, causing serious problems in public health and even death because of the failure of the antibiotic at therapeutic doses [10]. Furthermore, another problem for human health is the presence of antibiotic residues in animal-derived food, by the use of antibiotics for long periods of time, since it is associated in some cases with allergic reactions, imbalance of the intestinal microbiota, and especially, the development of antibacterial resistance [11].

Consequently, one of the measures taken in the face of the problems of bacterial resistance was the restriction of antibiotics at subtherapeutic doses in the EU in 2006 [12] and the USA in 2017 [13], and although in countries as Mexico they have not been officially banned, the Ministry of Agriculture and Rural Development (SADER), through its decentralized administrative body, the National Health Service, Food Safety and Food Quality (SENASICA), has promoted initiatives to prevent their use since 2012 [14, 15, 16, 17]. However, as a consequence of this measure, the incidence of enteric diseases in animals has increased significantly [18], as well as the use of antibiotics, but at therapeutic doses for the purpose of controlling and preventing diseases, which could lead to a worse scenario of bacterial resistance [2, 19, 20, 21]. In this context, the European One Health Action Plan against antimicrobial resistance calls for the phasing out of routine prophylactic (Prevent) and metaphylactic (Control) antimicrobial use in animal production and investment in the research of new alternatives [22], since they could be regulated in the coming years.

Therefore, the poultry industry has been under pressure to seek and investigate new alternatives to reduce the problems of bacterial resistance, prevent and control diseases, reduce the mortality rate, and finally promote the growth of animals. Among these alternatives, the most popular are probiotics (yeasts or bacteria) since it has been reported that they can improve the performance [23, 24], as well as prevent and control enteric pathogens in poultry [25, 26, 27]. Furthermore, it has been reported that probiotics could be an interesting alternative to prevent and control the toxic effects of aflatoxins. For these reasons, the probiotic market has expanded rapidly and is expected to grow to around 7% in 2020. However, this market is led mainly by Asia and Europe given the growing demand for dietary supplements [18].

Advertisement

2. Probiotics

Probiotics are defined as “live strains of strictly selected microorganisms which, when administered in adequate amounts, confer a health benefit on the host” [28]. The most common microorganisms used as probiotics in livestock production are lactic acid bacteria (LAB) from the genus Lactobacillus, Pediococcus, Lactococcus, Enterococcus, Streptococcus, and Leuconostoc. Nevertheless, only the genera Lactobacillus, Streptococcus, Pediococcus, Enterococcus, and Weissella are the most frequently used in poultry production [29]. Although the efficacy of probiotics reducing enteric pathogens is evident, one of the disadvantages is that they require refrigeration or lyophilization to survive for long storage periods or can be encapsulated to increase their stability/viability when included in the feed, which would increase the cost of production at the industrial level, making it unprofitable [30]. Unlike LAB, direct-fed microbials (DFM) as Bacillus spores, other types of probiotics, have several potential applications since they can be included as feed additives in poultry diets, due to their remarkable heat stability and long shelf life [31, 32]. Bacteria of the genus Bacillus are Gram-positive, frequently found in the soil. However, several studies have shown that Bacillus spores can also be present, germinate, and survive in the gastrointestinal tract (GIT) of different animal species [25]. The survival rate and persistence of some Bacillus strains in the GIT could be related to their capacity to synthesize biofilms, thus protecting themselves against the different conditions present in the gut [33]. Furthermore, another advantage of Bacillus strains is that they are frequently used by biotechnology companies for the production of enzymes and antibiotics. Therefore, these multifunctional microorganisms have different applications, since they are useful inside or outside a host [34, 35].

2.1 Mechanisms of action probiotics

2.1.1 Pathogenic bacteria

Although a large number of studies have shown the possible mechanisms by which probiotics have a beneficial action in inhibiting of pathogens, more studies are needed to elucidate them.

The possible modes of action of probiotics for the inhibition of pathogens include two basic mechanisms [29, 36, 37]: competitive exclusion and modulation of the host immune system (Figure 1). Competitive exclusion involves mechanisms such as (1) production of inhibitory compounds, that is, hydrogen peroxide, bacteriocins, and defensins [38, 39], (2) prevention of the pathogen adhesion [38], (3) competition for nutrients [40], and (4) reduction of toxin bioavailability [36]. Meanwhile, in the modulation of the host immune system, both innate and adaptive immune responses are involved [29]. The adaptive immune response depends on B and T lymphocytes to induce an antigen-specific response and produce antibodies [29, 41]. In contrast, physical and chemical barriers (innate immunity), such as intestinal epithelial cells (IEC), are the first line of defense to prevent the spread of pathogens and subsequent infections. Furthermore, IEC are the target cells for probiotics, which can improve the function of the intestinal barrier by stimulating the production of mucus and antimicrobial peptides such as defensins [42, 43].

Figure 1.

Mechanism of action of probiotics.

2.1.2 Aflatoxins

Similar as for pathogenic bacteria, probiotics can (1) compete for space and nutrients with aflatoxigenic mold strains, (2) degrade aflatoxins by the production of enzymes, or (3) avoid the intestinal absorption of AFB1 by its binding to the cell walls of probiotic strains [44].

Advertisement

3. Probiotic application in poultry industry

Although probiotics are considered potential alternatives to antibiotic use in poultry because they leave no residues in the meats and eggs given their modes of action, the variety of microorganisms in terms of species and even between strains of the same species, as well as their variation in metabolic activity, could affect their effectiveness. Furthermore, other factors that influence the effectiveness of probiotics in poultry are the species of origin, the probiotic preparation method, the survival of colonizing microorganisms in the gastrointestinal tract conditions, the environment where the birds are raised, the application time and administration route of probiotics, the immunologic state, the lineage of poultry, as well as age and concomitant use of antibiotics [45, 46]. Below are some of the applications of probiotics in poultry.

3.1 Effects of lactic acid bacteria against pathogens of importance in poultry

Several articles published by our laboratory have shown that the use of probiotics as a replacement of antibiotics in poultry production has had positive effects by reducing the growth of pathogens in in vitro models that simulate or not the three main compartments in birds (crop, proventriculus, and intestine) [47, 48], as well as the colonization of pathogens through the gastrointestinal tract in both turkeys and broiler chickens [26, 27, 49, 50, 51]. Although the results obtained have been promising, it is a fact that the isolated probiotics were characterized biochemically and by 16S rRNA sequence analyses (Microbial ID Inc., Newark, DE 19713, USA), subsequently, they were evaluated using in vitro models to determine their activity against pathogens, and, finally, the candidates were tested in in vivo models with the purpose of obtaining a well-characterized functional product.

Extensive research conducted by our laboratory determined the antimicrobial capability of several lactic acid bacteria (LAB) isolates mainly against Salmonella in in vitro models. However, only 11 were selected to produce a product called FloraMax®-B11 given their effect against Salmonella. Subsequently, these LAB were characterized by 16S rRNA sequence analyses (Table 1) [52].

LAB identification16S rRNA sequence analyses (Microbial ID Inc.)
18Pediococcus parvulus
24Weissella confusa
27Weissella confusa
29Pediococcus parvulus
36Lactobacillus salivarius
37BWeissella confusa
40Weissella confusa
44Weissella paramesenteroides
46Lactobacillus salivarius
48Lactobacillus salivarius
42Pediococcus parvulus

Table 1.

Identifications of FloraMax®-B11 (FM-B11) lactic acid bacteria (LAB).

However, since these LAB were grown together in a culture, the only LAB that remained viable were Lactobacillus salivarius and Pediococcus parvulus, two strains of poultry gastrointestinal origin. Despite this, in vitro studies showed that FloraMax®-B11 presented antimicrobial activity against Salmonella enteritidis, Escherichia coli (O157:H7), and Campylobacter jejuni [47] (Table 2). The antimicrobial activity of this probiotic culture could be due to the accumulation of primary metabolites such as lactic acid, ethanol, and carbon dioxide and to the production of other antimicrobial compounds such as bacteriocins [53]. Furthermore, the probiotic culture was capable of maintaining its viability under acidic conditions (pH = 3) for 4 h, which agrees with other studies where Lactobacillus spp. isolates were resistant to low pH, with high survival rates at pH 3.0 for 1 h [54]. Although probiotic bacteria need to survive passage through the stomach (pH 1.5–2.0) [55], and maintain their viability for 4 h or more [56] before reaching the intestine, the feed passage rate for birds is faster; therefore, bacterial acid tolerance is not as critical in chickens as it is in other animals [57]. Additionally, this probiotic culture grew at low and high temperatures for 4 h of incubation. However, the ability to grow at high temperatures is an important advantage since the production of lactic acid increases, and, therefore, the bacterial load decreases [58]. The probiotic culture was also able to tolerate high osmotic concentrations of NaCl, but it is extremely important since it has been reported that a high salt concentration could affect the physiology of probiotics, as well as their enzymatic activity, water activity, and metabolism [58]. Finally, this probiotic culture has its ability to tolerate bile salt concentrations of 0.4, 0.5, and 0.6% for 2, 4, and 24 h of incubation. Bile resistance of probiotics is related to their enzyme activity of bile salt hydrolase that helps to hydrolyze conjugated bile, reducing its toxic effect [59, 60].

Salmonella enteritidisEscherichia coli (O157:H7)Campylobacter jejuni
Lactobacillus salivarius+++
Pediococcus parvulus+++

Table 2.

In vitro assessment of antimicrobial activity of Lactobacillus salivarius and Pediococcus parvulus present in FloraMax®-B11 against enteropathogenic bacteria.

Symbols: +, inhibition.

Furthermore, the effect of this commercial product (FloraMax®-B11) has been evaluated in different models of infection both in broiler chickens and turkeys. In neonatal broilers, the administration of 1 × 106 cfu/bird FloraMax®-B11 by oral gavage 1 h after the chicks were challenged with Salmonella enteritidis (SE) and Salmonella typhimurium (ST) (1 × 104 cfu/bird) reduced the incidence of SE and ST, as well as the SE counts by >2.9 log, 24 h post-LAB administration [61] (Table 3). In contrast, there were no significant differences at 6- and 12-h post-LAB administration, but a slight reduction was observed at 12-h post-LAB administration. These data suggest that the mechanism to reduce Salmonella was initiated within the first 12 h after treatment. Probably the reduction of Salmonella is due to the set of mechanisms of action of probiotics: bacterial interactions (competitive exclusion) or stimulation of a host innate immune response. The competitive exclusion could have included competition for receptor sites, production of volatile fatty acids that are inhibitors of certain enteric pathogens, production of bacteriocins, or competition with pathogens and native flora for limiting nutrients [62]. Furthermore, since the Salmonella recovery was performed in the early stages of infection, the innate immune response could be responsible for the reduction of Salmonella.

Rep.TreatmentST cecal tonsil +/− (%)SE cecal tonsil +/− (%)Log SE cecal recovery (all samples)Log SE cecal recovery (only positive samples)
1Control20/25 (80)22/25 (88)3.81 ± 0.324.33 ± 0.17
LAB2/25 (8)*8/25 (32)*0.62 ± 0.19*1.95 ± 0.09*
2Control18/25 (72)25/25 (100)3.59 ± 0.233.59 ± 0.23
LAB2/25 (8)*7/25 (28)*0.42 ± 0.18*1.91 ± 0.29*
3Control20/25 (80)25/25 (100)3.91 ± 0.193.91 ± 0.19
LAB1/25 (4)*11/25 (40)*1.00 ± 0.25*2.22 ± 0.24*

Table 3.

Effect of lactic acid bacteria (LAB) on Salmonella typhimurium (ST) or Salmonella enteritidis (SE) recovered from cecal tonsils or ceca of broiler chicks 24-h post-LAB administration.

A significant (p ≤ 0.05) difference was observed between control and treated within a single experiment in each column.


In our other studies, the administration of FloraMax®-B11 in drinking water (106 cfu/mL) for 3 days post-SE challenge (104 cfu/bird) using two presentations, liquid and lyophilized significantly reduced the incidence of Salmonella [63], which agrees with other studies [64]. Furthermore, the administration of FloraMax®-B11 at the same concentration as the previous study after 1-h post-Salmonella Heidelberg (SH) challenge practically eliminated the concentration of SH, as well as its incidence, since only one sample was positive. However, in turkey poults under the same experimental conditions (Table 4), although similar significant results were observed at day 3 post-FloraMax®-B11 administration, it is clear that poults were more susceptible to SH colonization than chicks [51].

Treatment24 h72 h
Cecal tonsils1SH2 (log10 cfu/g of ceca content)Cecal tonsils1SH2 (log10 cfu/g of ceca content)
Control SH20/20 (100)7.04 ± 0.19a20/20 (100)6.05 ± 0.28a
FloraMax®-B1113/20 (65)*4.36 ± 0.74b9/20 (45)*2.15 ± 0.75b

Table 4.

In vivo evaluation of FloraMax-B11 against Salmonella Heidelberg (SH) at 24 and 72 h in poults.

Data expressed as positive/total poults (%).


Data expressed as mean ± SE.


p < 0.001.


a,bDifferent superscripts within columns indicate significant differences (p < 0.05).

Finally, trying to find FloraMax®-B11 applications in poultry, we opted for spray application since it could be more efficient and has lower cost than its application in drinking water since it is important to take into account water quality and medicator/proportioner function [65]. The results obtained were promising since when the probiotic was applied by spray and in drinking water, there was a reduction in the recovery of SE (55 and 50%, respectively; controls 85%) when chicks were held for 8 h prior to SE challenge and placement. In the same way, when probiotic was applied by spray or in drinking water and SE challenge occurred simultaneously, with placement 8 h after treatment, a marked and significant reduction of SE recovery was noted after 5d (10 and 40%, respectively; controls 55%). Furthermore, when the probiotic was sprayed and chickens were SE challenged simultaneously, with placement 8 h after treatment, a significant reduction of SE recovery was again noted in both the spray and DW application (80% controls, 15% spray, 15% drinking water) (Table 5). These results suggest that the spray application of this probiotic can be effective in protecting chicks against Salmonella infection. Furthermore, hatchery administration could prove to be a more effective way to administer probiotics because the chicks will be receiving the beneficial bacteria at the earliest possible time, in the absence of in ovo administration.

Treatment regimenGroupCecal tonsils
Exp. 1Exp. 2
Treat-challenge-place immediatelyControl95% (19/20)95% (19/20)
Probiotic (drinking water)75% (15/20)25% (5/25)**
Probiotic spray90% (18/20)80% (16/20)
Treat-hold 8 h-challenge-placeControl85% (17/20)70% (14/20)
Probiotic (drinking water)50% (10/20)*70% (14/20)
Probiotic spray55% (11/20)*80% (16/20)
Treat-challenge-hold 8 h-placeControl55% (11/20)80% (16/20)
Probiotic (drinking water)44% (7/20)*15% (2/20)*
Probiotic spray20% (2/20)**15% (2/20)*

Table 5.

Salmonella enteritidis recovery from cecal tonsils of broiler chicks 5-day post-challenge.

Indicates significant (p < 0.05) differences were observed between control and treated within a single experiment and treatment regime in each column.


Significantly (p < 0.01) different than all groups within a single experiment and treatment regime in each column.


In this regard, an in ovo study was performed to know the effectiveness of FloraMax®-B11 [66]. For this, 18-day-old embryos were candled and inoculated with either saline or 104 cfu FloraMax®-B11 via in ovo injection into the amnion. On day 21, chicks were pulled from hatchers to measure hatchability. Subsequently, all chickens were then orally gavaged with SE on the day of hatch (~104 cfu/chick) and maintained for 7 days. Salmonella recovery was done 24-h post-SE challenge. Body weight (BW) was determined at days 1, 3, and 7. In this experiment, a significant increase in BW was observed. Furthermore, chickens that received the probiotic culture showed a significant reduction in the incidence and counts of SE in cecal tonsils when compared with saline control chickens (Table 6).

TreatmentDay 1 BW (g)Day 3 BW (g)Day 7 BW (g)SE incidence cecal tonsils 24 h PILog SE/g of ceca content 24 h PI
Saline49.13 ± 0.30a62.53 ± 0.81b132.89 ± 3.06b20/20 (100%)7.13 ± 1.01a
FloraMax®-B1149.72 ± 0.36a65.42 ± 0.77a144.98 ± 3.02a9/20 (45%)*5.45 ± 1.25b

Table 6.

Evaluation of in ovo administration of FloraMax®-B11 on body weight and Salmonella enteritidis (SE) recovery in broiler chickens.

Indicates significant differences p < 0.001, n = 20/group.


a,bSuperscripts within columns indicate significant differences p < 0.05, n = 12/group.

These results agree with another study where the in ovo colonization with a probiotic could become an important method to reduce Salmonella and other intestinal bacterial infections in poultry [67]. Regarding the increase of BW in the group treated with the probiotic, this could be due to the significant morphometric changes in the duodenum and ileum observed at day 1 of age.

3.2 The use of direct-fed microbials (DFM) for the control of pathogens in poultry

Although the use of LAB has been promising for the control of pathogens such as Salmonella spp., as described above, it is important to mention that one limitation is their sensitivity to pelletizing processes for feed production (heating) [306869], environmental factors [70], and the low pH of the stomach and the presence of bile salts in the small intestine [71, 72]. For this reason, some strategies to increase the viability of these bacteria include their microencapsulation in polymer matrices [7374], as well as their freezing or lyophilization [75, 76]. However, production costs increase, so it becomes nonviable in animal production. Although LAB are better probiotics than Bacillus, the latter is more stable due to their ability to form spores, which are more resistant to severe environmental conditions, feed pelleting process with extreme temperatures, as well as tolerance to extremes of pH, dehydration, high pressures, and chemicals, and therefore, stability to long period storage conditions, making them suitable for commercialization [77, 78] since they could be used as direct-fed microbials (DFM) [68].

Previously in our laboratory, we have screened and identified Bacillus spp. isolates as DFM. Some of these demonstrated to be effective as potential DFM candidates by reducing Salmonella colonization and having a positive effect on the increase in body weight gained in both chickens and turkeys, as well as tolerance to acidic condition (pH = 2), high osmotic pressure (NaCl at 6.5%), and 0.037% bile salts after 24 h of incubation [79, 80, 81].

Several studies have reported that some Bacillus species are capable of producing different exogenous enzymes such as protease, lipase, cellulase, xylanase, phytase, and keratinase [82, 83, 84, 85, 86], which agrees with one of our studies already published [25]. These enzymes could improve the digestion of nutrients, making them more bioavailable, and also, they help to reduce intestinal viscosity in non-starch polysaccharide diets and decrease the substrates available for the growth of pathogenic bacteria. Considering this information, we performed a study in order to evaluate the effect of three Bacillus-DFM candidates with excellent to good relative enzyme activity values (cellulase and xylanase) on digesta viscosity and Clostridium perfringens (CP) proliferation in different poultry diets using an in vitro digestive model [87]. One of the three Bacillus strains was identified as Bacillus subtilis and the other two isolates as Bacillus amyloliquefaciens by 16S rRNA sequence analysis. Subsequently, Bacillus candidate strains were sporulated and mixed in equal amounts during the Bacillus-DFM preparation process [88] and incorporated into the experimental diets (108 spores/g). The results of this study demonstrated that Bacillus candidate significantly reduced the viscosity of non-corn-based diets. This could be due to the capability of these Bacillus strains to produce cellulase and xylanase, which could help improve the digestibility of cereals with high-soluble non-starch polysaccharides [89]. Furthermore, Bacillus-DFM candidate demonstrated effective antimicrobial properties against CP (Table 7), given their capability to produce antimicrobial-like compounds and/or compete for nutrients. Likewise, it was shown that the persistence of Bacillus-DFM candidate spores changes in each compartment of the in vitro digestive model mainly due to the conditions of pH and suggests that their full life cycle is developed in the gastrointestinal tract.

DietControl dietBacillus-DFM
Corn-based6.44 ± 0.19a6.68 ± 0.08a
Wheat-based7.12 ± 0.07a5.20 ± 0.18b
Barley-based7.50 ± 0.13a6.86 ± 0.11b
Rye-based7.15 ± 0.09a6.68 ± 0.12b
Oat-based6.96 ± 0.13a5.76 ± 0.07b

Table 7.

Concentration of Clostridium perfringens (CP)1 in different digested diets with or without inclusion of Bacillus-DFM candidate spore2.

Inoculum used 105 cfu of CP.


Data expressed in log10 cfu/mL.


a,bDifferent superscripts within a row indicate significant differences p < 0.05.

Based on the previous results, the effect of Bacillus-DFM candidate spores formed by an isolate of Bacillus subtilis and two of Bacillus amyloliquefaciens on growth performance, intestinal integrity, necrotic enteritis (NE) lesions, and ileal microbiota in broiler chickens using a previously established NE-challenged model [90] was evaluated [24]. This study consisted of three experimental groups: negative control (NC), positive control (PC), and Bacillus-DFM group (DFM). The last two groups were challenged with Salmonella typhimurium (ST, day 1), Eimeria maxima (EM, day 13), and Clostridium perfringens (CP, day 18–19). The overall results of performance showed that chickens supplemented with DFM had a significant body weight (BW) higher than PC. Furthermore, the body weight gain (BWG) and feed conversion ratio (FCR) were 59 g higher and 17 points lower, respectively, in the DFM group than PC (Table 8).

ItemNegative controlPositive controlDFM
BW, g/broiler
d 046.88 ± 0.64b46.54 ± 0.64b49.23 ± 0.68a
d 7127.14 ± 2.90a115.58 ± 3.27b123.05 ± 3.80ab
d 14273.80 ± 11.02b295.78 ± 12.10ab318.08 ± 13.57a
d 18457.79 ± 18.97ab456.32 ± 19.39b525.58 ± 17.92a
d 21603.81 ± 24.32a445.96 ± 18.50c507.77 ± 20.60b
BWG, g/broiler
d 0–780.39 ± 3.06a67.74 ± 3.24b75.08 ± 3.64ab
d 7–14147.01 ± 9.51b182.60 ± 9.48a196.22 ± 10.56a
d 14–18183.99 ± 9.85ab160.55 ± 9.02b198.31 ± 9.61a
d 14–21325.78 ± 15.58a152.13 ± 9.67b185.27 ± 10.52b
d 0–21552.72 ± 24.35a399.42 ± 19.79b458.58 ± 20.48b
FI, g/broiler
d 0–21808.21 ± 29.86a772.34 ± 10.66a805.21 ± 71.07a
FCR
d 0–211.46 ± 0.04b1.93 ± 0.10a1.76 ± 0.18ab

Table 8.

Evaluation of body weight (BW), body weight gain (BWG), feed intake (FI), and feed conversion ratio (FCR) in chickens supplemented with or without DFM on a necrotic enteritis challenge model1.

Data expressed as mean ± SE from 40 chickens (four replicates with 10 chicks each pen). p < 0.05.


a–cValues within columns with different superscripts differ significantly (p < 0.05).

This enhancement in the performance of chickens supplemented with Bacillus-DFM could be due to better digestibility of nutrients, maintenance of the beneficial gut microbiota, and promotion of a healthy intestinal integrity [48, 8791]. Furthermore, these results could relate to the low-serum FITC-d concentration, bacterial translocation (BT), ileal lesion (IL), and total intestinal IgA levels in the DFM group compared to the PC group given the low impact of EM and CP challenge since DFM could produce beneficial chemical compounds, has immunoregulatory capacity, and stimulates the homeostasis of the intestinal microbiota, resulting in a proper intestinal health status [92].

Microbiota analysis confirms that DFM played a vital role in restoring gut dysbiosis. Although only the phylum Proteobacteria was significantly lower in DFM group than PC group, it could be explained due to the antimicrobial properties of DFM against ST [25], a predisposing factor in the NE model. In contrast, the genus Lactobacillus was significantly predominant in both NC and DFM groups with respect to PC, but it was higher in the DFM group than NC group (Figure 2). It has been reported that DFM is capable of increasing the genus Lactobacillus, which plays a crucial role in preventing dysbiosis and maintaining gut integrity (homeostasis) [36, 93].

Figure 2.

Relative abundance of different phyla (A), families (B), and genera (C) in different treatment groups (NC, PC, and DFM). NA refers to those reads that were not assigned to the respective taxonomic levels.

Furthermore, Clostridium was significantly higher in PC group due to the change in the ileum microbiota caused by NE [94], whereas the genera Lactobacillus and Bacillus were more abundant in the DFM group, suggesting that these genera could alleviate the negative impacts caused by CP [95].

Finally, significant differences in beta diversity were found between NC versus PC and PC versus DFM (Figure 3), which agrees with another study where NE causes significant changes in the intestinal microbiota [96]. Interestingly, there was no difference in bacterial community structure between NC and DFM. It confirms again that DFM played a vital role in restoring the gut dysbiosis in this study.

Figure 3.

PCoA plot showing difference in microbial community structure between (A) NC and PC (ANOSIM; R = 0.40 and p < 0.05) and (B) DFM and PC (ANOSIM; R = 0.73 and p < 0.01).

3.3 The use of Bacillus-DFM candidate to prevent the toxic effects of aflatoxin B1 (AFB1) in poultry

Aflatoxin B1 (AFB1) is the predominant mycotoxin produced by several species of Aspergillus [97]. This mycotoxin has hepatotoxic and hepatocarcinogenic effects [98]. It has been reported that AFB1 has detrimental effects on performance parameters, which can cause serious economic problems in the poultry industry [99]. Therefore, the control of AFB1 is critical for producers. In this sense, the use of probiotics has proven effective in preventing and controlling the toxic effects of AFB1.

An in vitro study performed in our laboratory showed that 3 of 69 Bacillus spp. candidates were capable of biodegrading AFB1 since they reduced the fluorescence and area of clearance around each colony [100]. However, when these Bacillus spp. were tested in broiler chickens, no significant differences in performance parameters were observed when the groups were compared [101].

Despite the previous results, the Bacillus-DFM candidate spores formed by the isolate of Bacillus subtilis and the two of Bacillus amyloliquefaciens were included in the diets containing AFB1 to determine their effect on performance in broiler chickens fed with 2-ppm AFB1-contaminated diet [unpublished work from our laboratory]. The results are promising since the Bacillus-DFM improved performance of broilers, and even, there were no significant differences between the negative control (NC) and DFM group. It was due to the capacity of DFM to produce certain essential nutrients, extracellular enzymes, and growth factors to promote host growth [99, 102] (Table 9).

ItemNCAFB1DFMSEM2p-value
BW, g/broiler
d 046.23 ± 0.68a47.92 ± 0.72a48.12 ± 0.74a0.41740.1275
d 7133.29 ± 4.64a129.92 ± 2.78a137.02 ± 4.19a2.27630.4502
d 14320.92 ± 17.53a272.06 ± 8.54b318.42 ± 14.65a8.42150.0263
d 21640.10 ± 31.51a474.81 ± 15.57b571.60 ± 25.47a16.23610.0001
BWG, g/broiler
d 0–787.06 ± 4.24a82.00 ± 2.71a88.90 ± 4.15a2.17050.4103
d 7–14187.63 ± 13.82a142.13 ± 7.06b181.40 ± 11.38a6.73370.0097
d 14–21319.17 ± 16.08a202.75 ± 9.77c253.17 ± 14.89b9.5832<0.0001
d 0–21593.87 ± 31.21a426.88 ± 15.66c523.48 ± 25.42b16.21050.0001
FI, g/broiler
d 0–21750.55 ± 17.23a775.93 ± 3.51a731.97 ± 82.35a25.12920.8193
FCR
d 0–211.27 ± 0.06b1.82 ± 0.06a1.40 ± 0.06b0.08750.0016

Table 9.

Evaluation of body weight (BW), body weight gain (BWG), feed intake (FI), and feed conversion ratio (FCR) in broiler chickens consuming a corn-soybean-based diet contaminated with aflatoxin B1 (2 ppm) supplemented with or without DFM.

a–cSuperscripts within rows indicate significant difference at p < 0.05.

Advertisement

4. Conclusions

As it can be seen, probiotics could be considered a potential alternative to the use of antibiotics in poultry since it has been reported that they can improve the performance, as well as prevent and control enteric pathogens in poultry. However, their applications depend on the type of microorganism. In this regard, since lactic acid bacteria (LAB) are very sensitive to pelletizing processes for feed production (heating), environmental factors, and the low pH of the stomach, as well as the presence of bile salts in the small intestine, their administration in a single dose could be the most viable application especially to prevent bacterial diseases in both in ovo and broiler chickens. In contrast, Bacillus spp. direct-fed microbials (DFM) can be a better alternative since they are more stable because they can form spores. Therefore, DFM can be included in the feed, and, in addition, the production costs are lower than the microencapsulation and freezing or lyophilization processes that are used to maintain the viability of LAB. Finally, probiotics as Bacillus-DFM have also shown beneficial effects in preventing and controlling toxic effects of AFB1. Although the mechanisms by which the DFM reduce the effect of AFB1 are still known, our laboratory is working to elucidate the mechanism.

Advertisement

Acknowledgments

This research was supported by the Arkansas Biosciences Institute under the project: Development of an avian model for evaluation early enteric microbial colonization on the gastrointestinal tract and immune function. The authors thank the CONACyT for the doctoral scholarship number 270728.

References

  1. 1. Cheng G, Hao H, Xie S, Wang X, Dai M, Huang L, et al. Antibiotic alternatives: The substitution of antibiotics in animal husbandry? Frontiers in Microbiology. 2014;5:217
  2. 2. Hao H, Cheng G, Iqbal Z, Ai X, Hussain HI, Huang L, et al. Benefits and risks of antimicrobial use in food-producing animals. Frontiers in Microbiology. 2014;5:288
  3. 3. Van Boeckel TP, Brower C, Gilbert M, Grenfell BT, Levin SA, Robinson TP, et al. Global trends in antimicrobial use in food animals. Proceedings of the National Academy of Sciences. 2015;112(18):5649-5654
  4. 4. Van Boeckel TP, Glennon EE, Chen D, Gilbert M, Robinson TP, Grenfell BT, et al. Reducing antimicrobial use in food animals. Science. 2017;357(6358):1350-1352
  5. 5. Klein EY, Van Boeckel TP, Martinez EM, Pant S, Gandra S, Levin SA, et al. Global increase and geographic convergence in antibiotic consumption between 2000 and 2015. Proceedings of the National Academy of Sciences. 2018;115(15):E3463-E3470
  6. 6. Bartlett JG, Gilbert DN, Spellberg B. Seven ways to preserve the miracle of antibiotics. Clinical Infectious Diseases. 2013;56(10):1445-1450
  7. 7. Mehndiratta PL, Bhalla P. Use of antibiotics in animal agriculture & emergence of methicillin-resistant Staphylococcus aureus (MRSA) clones: Need to assess the impact on public health. The Indian Journal of Medical Research. 2014;140(3):339
  8. 8. Huyghebaert G, Ducatelle R, Van IF. An update on alternatives to antimicrobial growth promoters for broilers. The Veterinary Journal. 2011;187(2):182-188. Available from: http://www.sciencedirect.com/science/article/pii/S1090023310000869
  9. 9. Allen HK, Levine UY, Looft T, Bandrick M, Casey TA. Treatment, promotion, commotion: Antibiotic alternatives in food-producing animals. Trends in Microbiology. 2013;21(3):114-119
  10. 10. Zaman SB, Hussain MA, Nye R, Mehta V, Mamun KT, Hossain N. A review on antibiotic resistance: Alarm bells are ringing. Cureus. 2017;9(6):e1403. DOI: 10.7759/cureus.1403
  11. 11. Muaz K, Riaz M, Akhtar S, Park S, Ismail A. Antibiotic residues in chicken meat: Global prevalence, threats, and decontamination strategies: A review. Journal of Food Protection. 2018;81(4):619-627
  12. 12. EPC. Ban on Antibiotics as Growth Promoters in Animal Feed Enters into Effect. European Commission—IP/05/1687 [Internet]. 2005. Available from: http://europa.eu/rapid/press-release_IP-05-1687_en.htm
  13. 13. Editors A. U.S. Bans Antibiotics Use for Enhancing Growth in Livestock [Internet]. 2017. McGraw-Hill Education. Available from: https://www.accessscience.com:443/content/u-s-bans-antibiotics-use-for-enhancing-growth-in-livestock/BR0125171 OP—AccessScience
  14. 14. DOF. Acuerdo por el que se dan a conocer los Lineamientos para la Operación Orgánica de las actividades agropecuarias. México; 2013
  15. 15. SENASICA. Guía para la presentación e integración de los documentos para la autorización o registro de los productos y aditivos alimenticios para consumo por animales. 2015. Available from: http://analav.com.mx/wp-content/uploads/2016/08/Guía-regulación-ALIMENTICIOS-oct15.pdf
  16. 16. Zaidi MB, Dreser A, Figueroa IM. A collaborative initiative for the containment of antimicrobial resistance in Mexico. Zoonoses and Public Health. 2015;62:52-57
  17. 17. DOF. Acuerdo por el que se declara la obligatoriedad de la Estrategia Nacional de Acción contra la Resistencia a los Antimicrobianos. México; 2018
  18. 18. Park YH, Hamidon F, Rajangan C, Soh KP, Gan CY, Lim TS, et al. Application of probiotics for the production of safe and high-quality poultry meat. Korean Journal for Food Science of Animal Resources. 2016;36(5):567
  19. 19. Marshall BM, Levy SB. Food animals and antimicrobials: Impacts on human health. Clinical Microbiology Reviews. 2011;24(4):718-733
  20. 20. Borck Høg B, Korsgaard HB, Wolff Sönksen U, Bager F, Bortolaia V, Ellis-Iversen J, et al. DANMAP 2016-Use of Antimicrobial Agents and Occurrence of Antimicrobial Resistance in Bacteria from Food Animals, Food and Humans in Denmark [Internet]. 2016. Available from: http://orbit.dtu.dk/files/140535625/DANMAP_2016_LOW_241017.pdf
  21. 21. Founou LL, Founou RC, Essack SY. Antibiotic resistance in the food chain: A developing country-perspective. Frontiers in Microbiology. 2016;7:1881. Available from: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5120092/
  22. 22. European Parliament. European Parliament Resolution of 13 September 2018 on a European One Health Action Plan against Antimicrobial Resistance (AMR) (2017/2254(INI)). 2018; Available from: http://www.europarl.europa.eu/doceo/document/TA-8-2018-0354_EN.pdf
  23. 23. Latorre JD, Hernandez-Velasco X, Vicente JL, Wolfenden R, Hargis BM, Tellez G. Effects of the inclusion of a Bacillus direct-fed microbial on performance parameters, bone quality, recovered gut microflora, and intestinal morphology in broilers consuming a grower diet containing corn distillers dried grains with solubles. Poultry Science. 2017;96(8):2728-2735
  24. 24. Hernandez-Patlan D, Solis-Cruz B, Pontin KP, Hernandez X, Merino-Guzman R, Adhikari B, et al. Impact of a Bacillus direct-fed microbial on growth performance, intestinal barrier integrity, necrotic enteritis lesions and ileal microbiota in broiler chickens using a laboratory challenge model. Frontiers in Veterinary Science. 2019;6:108
  25. 25. Latorre JD, Hernandez-Velasco X, Wolfenden RE, Vicente JL, Wolfenden AD, Menconi A, et al. Evaluation and selection of Bacillus species based on enzyme production, antimicrobial activity, and biofilm synthesis as direct-fed microbial candidates for poultry. Frontiers in Veterinary Science. 2016;3:95
  26. 26. Yang Y, Latorre JD, Khatri B, Kwon YM, Kong BW, Teague KD, et al. Characterization and evaluation of lactic acid bacteria candidates for intestinal epithelial permeability and Salmonella typhimurium colonization in neonatal Turkey poults. Poultry Science. 2017;97(2):515-521
  27. 27. Prado-Rebolledo OF, Delgado-Machuca J d J, Macedo-Barragan RJ, Garcia-Márquez LJ, Morales-Barrera JE, Latorre JD, et al. Evaluation of a selected lactic acid bacteria-based probiotic on Salmonella enterica serovar enteritidis colonization and intestinal permeability in broiler chickens. Avian Pathology. 2017;46(1):90-94
  28. 28. Tsuda H, Miyamoto T. Guidelines for the evaluation of probiotics in food. Report of a joint FAO/WHO working group on drafting guidelines for the evaluation of probiotics in food guidelines for the evaluation of probiotics in food. Report of a joint FAO/WHO working group on drafting guidelines for the evaluation of probiotics in food, 2002. Food Science and Technology Research. 2010;16(1):87-92
  29. 29. Vieco-Saiz N, Belguesmia Y, Raspoet R, Auclair E, Gancel F, Kempf I, et al. Benefits and inputs from lactic acid bacteria and their bacteriocins as alternatives to antibiotic growth promoters during food-animal production. Frontiers in Microbiology. 2019;10:57
  30. 30. Tellez G, Pixley C, Wolfenden RE, Layton SL, Hargis BM. Probiotics/direct fed microbials for Salmonella control in poultry. Food Research International. 2012;45(2):628-633
  31. 31. Cartman ST, La Ragione RM, Woodward MJ. Bacillus subtilis spores germinate in the chicken gastrointestinal tract. Applied and Environmental Microbiology. 2008;74(16):5254-5258
  32. 32. Nguyen ATV, Nguyen DV, Tran MT, Nguyen LT, Nguyen AH, Phan T. Isolation and characterization of Bacillus subtilis CH 16 strain from chicken gastrointestinal tracts for use as a feed supplement to promote weight gain in broilers. Letters in Applied Microbiology. 2015;60(6):580-588
  33. 33. Barbosa TM, Serra CR, La Ragione RM, Woodward MJ, Henriques AO. Screening for Bacillus isolates in the broiler gastrointestinal tract. Applied and Environmental Microbiology. 2005;71(2):968-978
  34. 34. Priest FG. Extracellular enzyme synthesis in the genus Bacillus. Bacteriological Reviews. 1977;41(3):711
  35. 35. Azevedo EC, Rios EM, Fukushima K, Campos-Takaki GM. Bacitracin production by a new strain of Bacillus subtilis. Applied Biochemistry and Biotechnology. 1993;42(1):1
  36. 36. Caly DL, D’Inca R, Auclair E, Drider D. Alternatives to antibiotics to prevent necrotic enteritis in broiler chickens: A microbiologist’s perspective. Frontiers in Microbiology. 2015;6:1336
  37. 37. Alagawany M, El-Hack MEA, Farag MR, Sachan S, Karthik K, Dhama K. The use of probiotics as eco-friendly alternatives for antibiotics in poultry nutrition. Environmental Science and Pollution Research. 2018;25(11):10611-10618
  38. 38. Tiwari G, Tiwari R, Pandey S, Pandey P. Promising future of probiotics for human health: Current scenario. Chronicles of Young Scientists. 2012;3(1):17
  39. 39. Pan D, Yu Z. Intestinal microbiome of poultry and its interaction with host and diet. Gut Microbes. 2014;5(1):108-119
  40. 40. Kabir SML, Rahman MM, Rahman MB, Rahman MM, Ahmed SU. The dynamics of probiotics on growth performance and immune response in broilers. International Journal of Poultry Science. 2004;3(5):361-364
  41. 41. Khalighi A, Behdani R, Kouhestani S. Probiotics: A Comprehensive Review of their Classification, Mode of Action and Role in Human Nutrition. IntechOpen; 2016
  42. 42. Schlee M, Harder J, Köten B, Stange EF, Wehkamp J, Fellermann K. Probiotic lactobacilli and VSL# 3 induce enterocyte β-defensin 2. Clinical and Experimental Immunology. 2008;151(3):528-535
  43. 43. Bermudez-Brito M, Plaza-Díaz J, Muñoz-Quezada S, Gómez-Llorente C, Gil A. Probiotic mechanisms of action. Annals of Nutrition & Metabolism. 2012;61(2):160-174
  44. 44. Solis-Cruz B, Hernandez-Patlan D, Hargis B, Téllez G. Control of Aflatoxicosis in Poultry Using Probiotics and Polymers [Online First]. In: Micotoxins-Impact and management strategies. IntechOpen; 2018. DOI: 10.5772/intechopen.76371. Available from: https://www.intechopen.com/
  45. 45. Otutumi LK, De Moraes Garcia ER, Góis MB, Loddi MM. Variations on the efficacy of probiotics in poultry. In: Rigobelo EC, editor. Probiotic in Animals. Rijeka: InTech; 2012:203-230
  46. 46. Harimurti S, Hadisaputro W. Probiotics in Poultry. In: Beneficial microorganisms in agriculture, aquaculture and other areas. Cham: Springer; 2015:1-19
  47. 47. Menconi A, Kallapura G, Latorre JD, Morgan MJ, Pumford NR, Hargis BM, et al. Identification and characterization of lactic acid bacteria in a commercial probiotic culture. Bioscience of Microbiota, Food and Health. 2014;33(1):25-30
  48. 48. Latorre JD, Hernandez-Velasco X, Bielke LR, Vicente JL, Wolfenden R, Menconi A, et al. Evaluation of a Bacillus direct-fed microbial candidate on digesta viscosity, bacterial translocation, microbiota composition and bone mineralisation in broiler chickens fed on a rye-based diet. British Poultry Science. 2015;56(6):723-732
  49. 49. Higgins JP, Higgins SE, Wolfenden AD, Henderson SN, Torres-Rodriguez A, Vicente JL, et al. Effect of lactic acid bacteria probiotic culture treatment timing on Salmonella enteritidis in neonatal broilers. Poultry Science. 2010;89(2):243-247
  50. 50. Higgins SE, Wolfenden AD, Tellez G, Hargis BM, Porter TE. Transcriptional profiling of cecal gene expression in probiotic-and Salmonella-challenged neonatal chicks. Poultry Science. 2011;90(4):901-913
  51. 51. Menconi A, Wolfenden AD, Shivaramaiah S, Terraes JC, Urbano T, Kuttel J, et al. Effect of lactic acid bacteria probiotic culture for the treatment of Salmonella enterica serovar Heidelberg in neonatal broiler chickens and Turkey poults. Poultry Science. 2011;90(3):561-565
  52. 52. Tellez G, Higgins SE, Donoghue AM, Hargis BM. Digestive physiology and the role of microorganisms. Journal of Applied Poultry Research. 2006;15(1):136-144
  53. 53. Rattanachaikunsopon P, Phumkhachorn P. Lactic acid bacteria: Their antimicrobial compounds and their uses in food production. Annals of Biological Research. 2010;1(4):218-228
  54. 54. Fontana L, Bermudez-Brito M, Plaza-Diaz J, Munoz-Quezada S, Gil A. Sources, isolation, characterisation and evaluation of probiotics. The British Journal of Nutrition. 2013;109(S2):S35-S50
  55. 55. Dunne C, O’Mahony L, Murphy L, Thornton G, Morrissey D, O’Halloran S, et al. In vitro selection criteria for probiotic bacteria of human origin: Correlation with in vivo findings. The American Journal of Clinical Nutrition. 2001;73(2):386s-392s
  56. 56. Bakari D, Tatsadjieu NL, Mbawala A, Mbofung CM. Assessment of physiological properties of some lactic acid bacteria isolated from the intestine of chickens use as probiotics and antimicrobial agents against enteropathogenic bacteria. Innovative Romanian Food Biotechnology. 2011;8:33
  57. 57. Boonkumklao P, Kongthong P, Assavanig A. Acid and bile tolerance of Lactobacillus thermotolerans, a novel species isolated from chicken feces. Kasetsart Journal. 2006;40:13-17
  58. 58. Ibourahema C, Dauphin RD, Jacqueline D, Thonart P. Characterization of lactic acid bacteria isolated from poultry farms in Senegal. African Journal of Biotechnology. 2008;7(12):2006-2012
  59. 59. Du Toit M, Franz C, Dicks LMT, Schillinger U, Haberer P, Warlies B, et al. Characterisation and selection of probiotic lactobacilli for a preliminary minipig feeding trial and their effect on serum cholesterol levels, faeces pH and faeces moisture content. International Journal of Food Microbiology. 1998;40(1-2):93-104
  60. 60. Tanaka H, Doesburg K, Iwasaki T, Mierau I. Screening of lactic acid bacteria for bile salt hydrolase activity. Journal of Dairy Science. 1999;82(12):2530-2535
  61. 61. Higgins JP, Higgins SE, Vicente JL, Wolfenden AD, Tellez G, Bm H. Temporal effects of lactic acid bacteria probiotic culture on Salmonella in neonatal broilers. Poultry Science. 2007;86(8):1662-1666
  62. 62. Mead GC. Prospects for ‘competitive exclusion’ treatment to control Salmonella and other foodborne pathogens in poultry. Veterinary Journal. 2000;159(2):111-123
  63. 63. Vicente JL, Torres-Rodriguez A, Higgins SE, Pixley C, Tellez G, Donoghue AM, et al. Effect of a selected Lactobacillus spp.-based probiotic on Salmonella enterica serovar enteritidis-infected broiler chicks. Avian Diseases. 2008;52(1):143-146
  64. 64. Pascual M, Hugas M, Badiola JI, Monfort JM, Garriga M. Lactobacillus salivarius CTC2197 prevents Salmonella enteritidis colonization in chickens. Applied and Environmental Microbiology. 1999;65(11):4981-4986
  65. 65. Wolfenden AD, Pixley CM, Higgins JP, Higgins SE, Vicente J, Torres-Rodriguez A, et al. Evaluation of spray application of a Lactobacillus-based probiotic on Salmonella enteritidis colonization in broiler chickens. International Journal of Poultry Science. 2007;6(7):493-496
  66. 66. Teague KD, Graham LE, Dunn JR, Cheng HH, Anthony N, Latorre JD, et al. In ovo evaluation of FloraMax®-B11 on Marek’s disease HVT vaccine protective efficacy, hatchability, microbiota composition, morphometric analysis, and Salmonella enteritidis infection in broiler chickens. Poultry Science. 2017;96(7):2074-2082
  67. 67. De Oliveira JE, Van der Hoeven-Hangoor E, Van de Linde IB, Montijn RC, Van Der Vossen J. In ovo inoculation of chicken embryos with probiotic bacteria and its effect on posthatch Salmonella susceptibility. Poultry Science. 2014;93(4):818-829
  68. 68. Tellez G, Rodríguez-Fragoso L, Kuttappan VA, Kallapura G, Velasco X, Menconi A, et al. Probiotics for human and poultry use in the control of gastrointestinal disease: A review of real-world experiences. Alternative and Integrative Medicine. 2013;2:1-6
  69. 69. Guillermo T, Andrea L, Juan DL, Xochitl H-V, Billy MH, Todd C. Food-producing animals and their health in relation to human health. Microbial Ecology in Health and Disease. 2015;26(1):25876
  70. 70. Praepanitchai O-A, Noomhorm A, Anal AK. Survival and behavior of encapsulated probiotics (Lactobacillus plantarum) in calcium-alginate-soy protein isolate-based hydrogel beads in different processing conditions (pH and temperature) and in pasteurized mango juice. BioMed Research International. 2019;2019:1-8
  71. 71. Iravani S, Korbekandi H, Mirmohammadi SV. Technology and potential applications of probiotic encapsulation in fermented milk products. Journal of Food Science and Technology. 2015;52(8):4679-4696
  72. 72. Samedi L, Charles AL. Viability of 4 probiotic bacteria microencapsulated with arrowroot starch in the simulated gastrointestinal tract (GIT) and yoghurt. Food. 2019;8(5):175
  73. 73. Gbassi GK, Vandamme T. Probiotic encapsulation technology: From microencapsulation to release into the gut. Pharmaceutics. 2012;4(1):149-163
  74. 74. Evivie SE, Huo G-C, Igene JO, Bian X. Some current applications, limitations and future perspectives of lactic acid bacteria as probiotics. Food & Nutrition Research. 2017;61(1):1318034
  75. 75. Chávarri M, Marañón I, Villarán MC. Encapsulation technology to protect probiotic bacteria. In: Probiotics. IntechOpen; 2012
  76. 76. Montel Mendoza G, Pasteris SE, Otero MC, Fatima Nader-Macías ME. Survival and beneficial properties of lactic acid bacteria from raniculture subjected to freeze-drying and storage. Journal of Applied Microbiology. 2014;116(1):157-166
  77. 77. Vreeland RH, Rosenzweig WD, Powers DW. Isolation of a 250 million-year-old halotolerant bacterium from a primary salt crystal. Nature. 2000;407(6806):897
  78. 78. Cartman ST, La Ragione RM, Woodward MJ. Bacterial spore formers as probiotics for poultry. Food Science & Technology Bulletin Functional Foods. 2007;4(217):30
  79. 79. Shivaramaiah S, Pumford NR, Morgan MJ, Wolfenden RE, Wolfenden AD, Torres-Rodriguez A, et al. Evaluation of Bacillus species as potential candidates for direct-fed microbials in commercial poultry. Poultry Science. 2011;90(7):1574-1580
  80. 80. Wolfenden RE, Pumford NR, Morgan MJ, Shivaramaiah S, Wolfenden AD, Pixley CM, et al. Evaluation of selected direct-fed microbial candidates on live performance and Salmonella reduction in commercial Turkey brooding houses. Poultry Science. 2011;90(11):2627-2631
  81. 81. Menconi A, Morgan MJ, Pumford NR, Hargis BM, Tellez G. Physiological properties and Salmonella growth inhibition of probiotic Bacillus strains isolated from environmental and poultry sources. International Journal of Bacteriology. 2013;2013:1-8
  82. 82. Monisha R, Uma MV, Murthy VK. Partial purification and characterization of Bacillus pumilus xylanase from soil source. Kathmandu University Journal of Science, Engineering and Technology. 2009;5:137-148
  83. 83. Mazotto AM, Coelho RRR, Cedrola SML, de Lima MF, Couri S, Paraguai de Souza E, et al. Keratinase production by three Bacillus spp. using feather meal and whole feather as substrate in a submerged fermentation. Enzyme Research. 2011;2011:1-7
  84. 84. Arpana M, Gulab S, Varsha G, Anita Y, Aneja KR, Gautam SK, et al. Isolation and biochemical characterization of acido-thermophilic extracellular phytase producing bacterial strain for potential application in poultry feed. Jundishapur Journal of Microbiology. 2011;4(4):273-282
  85. 85. Shah K. Purification and characterization of lipase from B. subtilis Pa2. Journal of Biochemical Technology. 2012;3(3):292-295
  86. 86. Jani SA, Chudasama CJ, Patel DB, Bhatt PS, Patel HN. Optimization of extracellular protease production from alkali thermo tolerant actinomycetes: Saccharomonospora viridis SJ-21. Bulletin of Environment, Pharmacology and Life Sciences. 2012;1(6):84-92
  87. 87. Latorre JD, Hernandez-Velasco X, Kuttappan VA, Wolfenden RE, Vicente JL, Wolfenden AD, et al. Selection of Bacillus spp. for cellulase and xylanase production as direct-fed microbials to reduce digesta viscosity and Clostridium perfringens proliferation using an in vitro digestive model in different poultry diets. Frontiers in Veterinary Science. 2015;2:25
  88. 88. Zhao S, Deng L, Hu N, Zhao B, Liang Y. Cost-effective production of Bacillus licheniformis using simple netting bag solid bioreactor. World Journal of Microbiology and Biotechnology. 2008;24(12):2859-2863
  89. 89. Wang ZR, Qiao SY, Lu WQ , Li DF. Effects of enzyme supplementation on performance, nutrient digestibility, gastrointestinal morphology, and volatile fatty acid profiles in the hindgut of broilers fed wheat-based diets. Poultry Science. 2005;84(6):875-881
  90. 90. Shivaramaiah S, Wolfenden RE, Barta JR, Morgan MJ, Wolfenden AD, Hargis BM, et al. The role of an early Salmonella typhimurium infection as a predisposing factor for necrotic enteritis in a laboratory challenge model. Avian Diseases. 2011;55(2):319-323
  91. 91. Zhou M, Zeng D, Ni X, Tu T, Yin Z, Pan K, et al. Effects of Bacillus licheniformis on the growth performance and expression of lipid metabolism-related genes in broiler chickens challenged with Clostridium perfringens-induced necrotic enteritis. Lipids in Health and Disease. 2016;15(1):48
  92. 92. Jadamus A, Vahjen W, Simon O. Growth behaviour of a spore forming probiotic strain in the gastrointestinal tract of broiler chicken and piglets. Archives of Animal Nutrition. 2001;54(1):1-17
  93. 93. Jin LZ, Ho YW, Abdullah N, Jalaludin S. Probiotics in poultry: Modes of action. World’s Poultry Science Journal. 1997;53(4):351-368
  94. 94. Prescott JF, Smyth JA, Shojadoost B, Vince A. Experimental reproduction of necrotic enteritis in chickens: A review. Avian Pathology. 2016;45(3):317-322
  95. 95. Thibodeau A, Fravalo P, Yergeau É, Arsenault J, Lahaye L, Letellier A. Chicken caecal microbiome modifications induced by Campylobacter jejuni colonization and by a non-antibiotic feed additive. PLoS One. 2015;10(7):e0131978
  96. 96. Stanley D, Wu S-B, Rodgers N, Swick RA, Moore RJ. Differential responses of cecal microbiota to fishmeal, Eimeria and Clostridium perfringens in a necrotic enteritis challenge model in chickens. PLoS One. 2014;9(8):e104739
  97. 97. Wilson KA, Kung RW, Wetmore SD. Toxicology of DNA adducts formed upon human exposure to carcinogens: Insights gained from molecular modeling. In: Advances in Molecular Toxicology. Elsevier; 2016;10:293-360
  98. 98. Rawal S, Kim JE, Coulombe R Jr. Aflatoxin B1 in poultry: Toxicology, metabolism and prevention. Research in Veterinary Science. 2010;89(3):325-331
  99. 99. Oguz H, Kurtoglu V. Effect of clinoptilolite on performance of broiler chickens during experimental aflatoxicosis. British Poultry Science. 2000;41(4):512-517
  100. 100. Galarza-Seeber R, Latorre JD, Hernandez-Velasco X, Wolfenden AD, Bielke LR, Menconi A, et al. Isolation, screening and identification of Bacillus spp. as direct-fed microbial candidates for aflatoxin B1 biodegradation. Asian Pacific Journal of Tropical Biomedicine. 2015;5(9):702-706
  101. 101. Galarza-Seeber R, Latorre JD, Wolfenden AD, Hernandez-Velasco X, Merino-Guzman R, Ledoux DR, et al. Evaluation of Bacillus spp. as direct fed microbial (DFM) candidates for aflatoxin B1 biodegradation in broiler chickens. International Journal of Probiotics and Prebiotics. 2016;11(1):29-37
  102. 102. Guo M, Hao G, Wang B, Li N, Li R, Wei L, et al. Dietary administration of Bacillus subtilis enhances growth performance, immune response and disease resistance in Cherry Valley ducks. Frontiers in Microbiology. 2016;7:1975

Written By

Daniel Hernandez-Patlan, Bruno Solis-Cruz, Billy M. Hargis and Guillermo Tellez

Submitted: 04 May 2019 Reviewed: 25 July 2019 Published: 11 September 2019