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To evaluate the effects of dietary protein levels and probiotic supplementation on microbial intestinal fermentation, diarrhea incidence, and performance in weaned piglets, 162 piglets were randomly assigned to three treatments: high-protein diet (20%), with antibiotics (HPa); high-protein diet (20%), without antibiotics (HP); and low-protein diet, without antibiotics but with probiotics (LPpb). Piglets and feed were weighed weekly to calculate the average daily gain (ADG), average daily feed intake (ADFI), and gain: feed ratio (G:F). Four piglets per treatment were killed on day 21 postweaning to collect ileal and colon digesta for measurement of short-chain fatty acids (SCFAs), branched-chain fatty acids (BCFAs), lactic acid (LA), and ammonia (AM). In the digesta collected from the ileum and colon, piglets fed the HPa diet had the lowest concentration of volatile fatty acids. Additionally, the HP diet produced the highest concentration of BCFAs, while the LPpb diet produced moreacetic, propionic, and butyric acids than the HPa and HP diets. Piglets fed the HP diet had higher incidence and severity of diarrhea than piglets fed LPpb and HPa diets, and similar values were observed between these two groups. The second week postweaning was the most critical for diarrhea measurements; during the second week, animals had higher incidence and severity of diarrhea. Piglets fed the HPa and LPpb diets had similar ADGs, while those fed the HP diet had the poorest ADG. Similar results were observed with ADFI and G:F. A low-protein diet supplemented with probiotics changed the fermentation profile, reducing toxic metabolites, promoting gut health, decreasing the incidence and severity of postweaning diarrhea, and improving the performance of piglets.
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Food and Nutrition Sciences, 2014, 5, **-**
Published Online September 2014 in SciRes. http://www.scirp.org/journal/fns
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How to cite this paper: Author 1, Author 2 and Author 3 (2014) Paper Title. Food and Nutrition Sciences, 5, **-**.
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Microbial Fermentation Patterns, Diarrhea
Incidence, and Performance in Weaned
Piglets Fed a Low Protein Diet
Supplemented with Probiotics
Konisgmar Escobar García1, Tércia Cesária Reis de Souza1*, Gerardo Mariscal Landín2,
Araceli Aguilera Barreyro1, María Guadalupe Bernal Santos1, José Guadalupe Gómez Soto1
1Doctorado en Ciencias Biológicas, Facultad de Ciencias Naturales, Universidad Autónoma de Querétaro,
Querétaro, México
2CENID-Fisiología, Instituto Nacional de Investigación Forestal Agrícola y Pecuaria, Colón, México
Email: *tercia@uaq.mx
Received **** 2014
Copyright © 2014 by authors and Scientific Research Publishing Inc.
This work is licensed under the Creative Commons Attribution International License (CC BY).
http://creativecommons.org/licenses/by/4.0/
Abstract
To evaluate the effects of dietary protein levels and probiotic supplementation on microbial intes-
tinal fermentation, diarrhea incidence, and performance in weaned piglets, 162 piglets were ran-
domly assigned to three treatments: high-protein diet (20%), with antibiotics (HPa); high-protein
diet (20%), without antibiotics (HP); and low-protein diet, without antibiotics but with probiotics
(LPpb). Piglets and feed were weighed weekly to calculate the average daily gain (ADG), average
daily feed intake (ADFI), and gain: feed ratio (G:F). Four piglets per treatment were killed on day
21 postweaning to collect ileal and colon digesta for measurement of short-chain fatty acids
(SCFAs), branched-chain fatty acids (BCFAs), lactic acid (LA), and ammonia (AM). In the digesta
collected from the ileum and colon, piglets fed the HPa diet had the lowest concentration of vola-
tile fatty acids. Additionally, the HP diet produced the highest concentration of BCFAs, while the
LPpb diet produced moreacetic, propionic, and butyric acids than the HPa and HP diets. Piglets fed
the HP diet had higher incidence and severity of diarrhea than piglets fed LPpb and HPa diets, and
similar values were observed between these two groups. The second week postweaning was the
most critical for diarrhea measurements; during the second week, animals had higher incidence
and severity of diarrhea. Piglets fed the HPa and LPpb diets had similar ADGs, while those fed the
HP diet had the poorest ADG. Similar results were observed with ADFI and G:F. A low-protein diet
supplemented with probiotics changed the fermentation profile, reducing toxic metabolites, pro-
moting gut health, decreasing the incidence and severity of postweaning diarrhea, and improving
the performance of piglets.
*
Corresponding author.
K. E. García et al.
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Keywords
Diarrhea, Intestinal Fermentation, Piglet, Probiotics, SCFAs
1. Introduction
The use of vegetable protein sources in diets offered at weaning, when newly weaned piglets have a limited di-
gestive capacity, promotes fermentation of undigested protein by opportunistic microorganisms, producing
branched-chain fatty acids (BCFAs) and ammonia (AM). BCFAs and ammonia are toxic metabolites for the in-
testinal mucosa and most likely trigger of postweaning diarrhea and poor performance in piglets [1] [2]. Thus,
postweaning diarrhea is associated with the consumption of high-protein diets [3] [4].
To prevent postweaning diarrhea, piglets are often given antibiotics; however, antibiotics have been banned
for use in livestock for human consumption, exacerbating postweaning diarrhea and related problems in piglets.
Consequently, research has focused on looking for alternatives to replace antibiotics in piglet diets. One alterna-
tive is the use of low-protein diets: it has been hypothesized that consumption of a low crude protein diet reduc-
es the availability of substrates for bacterial fermentation [5] [6] and improves fecal consistency [5] [7]-[9],
which have direct effects on postweaning diarrhea [3] [9] [10]. Another alternative is the use of probiotics,
which have shown to have beneficial effects on intestinal health and productive performance in newly weaned
piglets [10] [11]. While previous studies have evaluated separately the use of low-protein diets and diets includ-
ing probiotics with different responses, no studies have assessed the effects of combining both treatments.
The aim of this study was to evaluate the effects of a combination of a low protein diet with probiotics on the
fermentation patterns of the ileum and colon, the incidence and severity of diarrhea, and piglet performance
during the first three weeks postweaning. Additionally, this diet was compared toa high protein diet with or
without antibiotics.
2. Materials and Methods
The experiment was conducted at the experimental farm of CENID-Physiology (INIFAP, Mexico). The protocol
was reviewed and approved by the bioethical committee of the Faculty of Natural Sciences of Autonomous
University of Queretaro. The experimental animals were treated according to the guidelines of the Mexican offi-
cial norm (NOM-062-ZOO-1999) for production, care, and use of animals for experimentation [12] and the
guidelines of the International Guiding Principles for Biomedical Research Involving Animals [13].
2.1. Animals and Diets
One hundred sixty-two (Fertilis 20 ×G Performance, Genetiporc) piglets were used. Animals were weaned at 20
± 1.4 days with an average body weight of 6.6 ± 1.1 kg. Piglets were housed in groups of six per pen, with nine
pens per treatment. Experimental treatments (diets) were as follows: high-protein diet (20%) with antibiotics
(HPa); high-protein diet (20%) without antibiotics (HP); and low-protein diet (16%) without antibiotics and with
probiotics (LPpb), as described in Table 1. High-protein diets provided amino acid requirements for this pro-
duction phase [14]; lysine, methionine, threonine, tryptophan, and valine were included in the low-protein diet to
cover nutritional requirements of the piglets [14]. The probiotic used in the LPpb group included Bacillus subti-
lis and B. licheniformis (1:1).
2.2. Animal Management, Performance Data, and Fecal Scores
Piglets were placed in a temperature-controlled room (30˚C, 28˚C, and 26˚C ± 2˚C during the first, second, and
third weeks postweaning, respectively). Animals were housed in weaning pens, with a nipple drinker and a
feeder with six spaces; piglets had free access to feed and water throughout the entire experimental period. Pig-
lets were weighed at the beginning of the experimental period and then every week thereafter. At the end of each
week, the feed intake of each pen was measured by determining the difference between the amount of feed of-
fered and the amount of feed rejected. These values were used to estimate feed efficiency (gain: feed ratio, G:F).
To calculate the average daily feed intake (ADFI) and the average daily gain (ADG), the weekly measurements
K. E. García et al.
3
Table 1. Ingredients and chemical compositions of experimental diets.
Treatments
HPa HP LPpb
INGREDIENTS (%)
Maize 43.57 43.62 55.67
Soybean meal 15.00 15.00 4.02
Fish Menhaden meal 1.00 1.00 1.00
Soybean protein concentrate 10.49 10.49 8.73
Whey dried 24.69 24.69 24.69
Maize oil 2.00 2.00 2.00
Lysine 0.22 0.22 0.50
Aminogut* 0.80 0.80 0.80
Threonine 0.02 0.02 0.15
Methionine 0.04 0.04 0.08
Tryptophan 0.01 0.01 0.08
Valine -- -- 0.17
Salt 0.01 0.01 0.01
Sodium carbonate 0.89 0.89 0.73
Limestone 0.71 0.71 0.82
LincoSpectin Premix** 0.05 -- --
Vitamins and minerals***** 0.30 0.30 0.30
Gustor*** 0.20 0.20 0.20
Bioplus 2B**** -- -- 0.05
CHEMICAL COMPOSITION
Dry matter (%) 89.7 91.7 90.3
Crude protein (%) 20.0 19.9 16.1
Ash (%) 5.9 5.8 5.6
Crude fat (%) 3.4 3.6 1.8
NDF (%) 5.8 5.0 5.6
ME (kcalkg1) 3300 3300 3300
HPa: high-protein diet with antibiotics; HP: high-protein diet without antibiotics; LPpb: low-protein diet without antibiotics and with
probiotics; *Aminogut, L-glutamine and L-glutamic acid (1:1), (Ajinomoto, Japan); **Linco Spectin premix, 2.2g lincomycin, 2.2 g spec-
tinomycin (Zoetis, USA); ***Gustorsodium butyrate, (Norel, Spain); ****Bioplus 2B: B. subtilis and B. licheniformis (1:1; Crh Hansen,
Denmark);*****Vitamin and mineral premix: (vitamins per kg of diet: vitamin A 10,200 IU; vitamin D 1,980 IU; vitamin E 60 IU; vita-
min K 1.20 mg; choline 967 mg; niacin 36 mg; pantothenate 17 mg; riboflavin 7.2 mg; vitamin B12 38 μg; thiamine 0.30 mg; pyridoxine
0.31 mg; biotin 0.08 mg; folate 0.75 mg mineral/kg diet; copper 14.4 mg; iodine 800 mg; iron 105 mg; manganese 36 mg; selenium 0.3
mg; zinc 144 mg).
were divided by seven (days of a week). Incidence of diarrhea was measured daily in each pen by direct obser-
vation by two different evaluators. The severity of diarrhea was measured through visual evaluation of the fecal
consistency, using a score from 0 to 3, where 0 indicated normal feces, 1 indicated mild diarrhea, 2indicated
moderate diarrhea, and 3 indicated severe diarrhea. The daily score of each pen was averaged every week to
calculate the severity of diarrhea [3].
K. E. García et al.
4
2.3. Sampling and Analysis
At day 21 after weaning, four animals from each group (one per experimental unit [pen]) were stunned using
CO2 and euthanized by exsanguination by severing the jugular vein. A midline incision was made in the abdo-
men to expose the digestive tract. The small and large intestines were removed from the abdominal cavity to
collect the terminal ileum and proximal colon digesta, which were immediately frozen in liquid nitrogen and
stored at 80˚C until analysis. Concentrations of acetic, propionic, butyric, and valeric acids (SCFAs); isobutyr-
ic, isovaleric, and isocaproic acids (BCFAs) [15] and lactic acid (LA) [16] were measured by gas chromatogra-
phy; and ammonia (AM) by distillation [17]. Diets were analyzed by Association of Official Analytical Chem-
ists methods [17]: moisture (934.01), ashes (900.02), crude protein (984.13), crude fat (954.16). Neutral Deter-
gent Fiber (NDF) were measured by Van Soest method [18].
2.4. Statistical Analysis
Fermentation profiles were analyzed using a completely randomized design, considering the piglet as the expe-
rimental unit. ADG, ADFI, and G:F were analyzed using a completely randomized design, considering the pen
as the experimental unit. In both cases, α-value of 0.05 was used to assess significance, and means were com-
pared by Tukey’s test using the GLM procedure of SAS [19] [20]. The presence and severity of diarrhea were
analyzed as repeated measures on a time design, considering the pen as the experimental unit using the Proc Mix
procedure of SAS [20].
3. Results
3.1. Microbial Fermentation End Products
Fermentation profiles in the ileum and colon were affected by diet (Table 2). Piglets fed HPa had lower concen-
tration of total VFAs (Volatile Fatty Acids) (P <0.001) than piglets fed diets without antibiotics (HP and LPpb).
The effects of antibiotic on the VFAs profile in the ileum and colon were different. The absence of antibiotic in
the HP diet increased SCFA production in the ileum, but decreased SCFA production in the colon (P < 0.001).
Animals fed the LPpb diet had higher concentration of SCFAs in the ileum and colon (P < 0.001) than animals
fed the HP or HPa diet. Acetate was the most abundant SCFAs in piglets fed the LPpb diet, followed by butyrate
and propionate. Higher concentrations of BCFAs were observed in the digesta, in both ileum and colon, of ani-
mals fed the HP diet than in those fed the HPa and LPpb diets (P < 0.001). LA concentrations in the ileum and
colon were higher in piglets fed the LPpb diet (P < 0.001) than in those fed the HP and HPa diets. AM concen-
trations were higher in animals fed the HP diet (P < 0.001) than in those receiving the HPa and LPpb diets (Ta-
ble 2).
3.2. Postweaning Diarrhea
All animals had mild diarrhea throughout the experimental period (Figure 1). However, animals fed the HP diet
had the highest incidence and severity of diarrhea (P < 0.01), while the other two groups (HPa and LPpb) exhi-
bited similar incidence and severity. The highest incidence and severity of diarrhea were observed during the
second week postweaning (P < 0.01). Both, incidence and severity of diarrhea, decreased during the third week
postweaning (P < 0.01), returning to values similar to those observed during the first week postweaning.
3.3. Performance
Piglets fed the HPa and LPpb diets showed similar performance parameters (P > 0.05) for all weeks and for the
entire experimental period (Figure 2). In contrast, animals fed the HP diet had significantly poorer performance
than the other two groups of animals (P < 0.05), except for ADFI during the first week postweaning and G:F
during the second week postweaning.
4. Discussion
4.1. Fermentation Pattern
The presence of antibiotics and level of dietary protein in the diet modify the fermentative activity of the intes-
K. E. García et al.
5
Table 2. Volatile fatty acids, lactic acid (μmol/g), and ammonia (mg/kg) production in the ileum and colon.
Treatments P SEM
HPa HP LPpb
Ileum
Acetate 11c 15b 82a *** 0.07
Butyrate 6.3c 7.3b 36a *** 0.06
Propionate 5.2b 4.3c 27a *** 0.06
Valerate 7.2b 19a 2.1c *** 0.04
Isobutyrate 2.5b 32a 2.2c *** 0.06
Isovalerate 3.7b 21a 1.9c *** 0.06
Isocaproate 2.2b 15a 0.7c *** 0.03
VFAs 38c 116b 152a *** 0.28
SCFAs 29c 46b 147a *** 0.19
BCFAs 8.5b 69a 4.8c *** 0.13
Lactate 16c 30b 41a *** 0.20
Ammonia 362b 514a 190c *** 1.9
Colon
Acetate 31b 19c 121a *** 0.06
Butyrate 12b 11c 46a *** 0.09
Propionate 11b 9.4c 38a *** 0.07
Valerate 15a 9.3b 1.8c *** 0.01
Isobutyrate 7.5b 58a 2.9c *** 0.02
Isovalerate 9.3b 36a 2.5c *** 0.04
Isocaproate 10b 17a 1.3c *** 0.01
VFAs 97c 162b 214a *** 0.16
SCFAs 70b 49c 207a *** 0.12
BCFAs 27b 112a 6.5c *** 0.05
Lactate 1.2c 1.9b 2.3a *** 0.008
Ammonia 400b 1077 275c *** 4.5
HPa: high-protein diet with antibiotics; HP: high-protein diet without antibiotics; LPpb: low-protein diet without antibiotics and with
probiotics; VFAs: volatile fatty acids; SCFAs: short-chain fatty acids; BCFAs: branched-chain fatty acids; ***P < 0.001; SEM: standard
error of the mean; abcdifferent letters in the same row show statistical differences.
tinal flora, as demonstrated by the results of the HP and LPpb groups in the present study. Animals in these
groups that did not consume antibiotics showed the highest concentrations of the most microbial fermentation
products (VFAs). In both, the ileum and colon of these animals, the quantity of VFAs was influenced by the
concentration of SCFAs. This was supported by the observation that piglets fed the LPpb diet had higher con-
centration of SCFAs than animals of the other two groups (HPa and HP) (P < 0.01), while piglets fed the HP di-
et had higher concentration of BCFAs than the other two groups (HPa and LPpb) (P < 0.01). Animals fed the
LPpb diet exhibited similar profiles of SCFAs in the colon and in the ileum. However, animals fed the HP diet
exhibited lower concentrations of SCFAs than those fed the HPa diet (with antibiotics).
K. E. García et al.
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Figure 1. Effect of treatments and postweaning weeks on the in-
cidence and severity of diarrhea. HPa: high-protein diet with anti-
biotics; HP: high-protein diet without antibiotics; LPpb: low-pro-
tein diet without antibiotics and with probiotics; WK1: first week
postweaning; WK2: second week postweaning; WK3: third week
postweaning; abcbars with different letters show statistical differ-
ences (P < 0.01).
These data indicate that antibiotics reduced overall fermentation in the ileum, but both antibiotics and protein
levels affected intestinal fermentation in the colon. Indeed, the absence of antibiotics and presence of a large
quantity of undigested protein in the hindgut probably favored the growth of microorganisms capable of fer-
menting protein (BCFAs and AM producers), which could limit the growth of other types of microorganisms
(generally SCFAs producers). These results were consistent with those of several other studies [1]-[3] [9] in
which the use of antibiotics showed to affect intestinal fermentation.
Microbial fermentation of protein increases AM production and fermentation of branched amino acids, gene-
rating BCFAs [2]. Thus, the level of dietary protein is a key factor that modifies the intestinal microenvironment.
This idea was supported by the high concentrations of BCFAs and AM in the gut of animals fed the HP diet in
the present study. This may be because piglets do not digest dietary protein well, and undigested protein re-
maining in the intestinal lumen can serve as a substrate for bacterial fermentation. BCFAs and AM are harmful
metabolites to intestinal mucosa because they are able to alter the acid-base balance, an essential factor for water
absorption in the hindgut, and this probably one of the triggers postweaning diarrhea [1] [21].
Conversely, when the protein level was reduced and probiotics were added (LPpb diet), SCFAs and LA pro-
duction were increased. This probably resulted from differences in the quantity of major ingredients of the diet
required to reduce the protein level; in the LPpb diet, protein sources were reduced, while corn was increased
(Table 1), providing a greater amount of starch, which is an important substrate to amylolytic bacteria, resulting
in an increased production of acetic and propionic acids. In turn, acetic acid can be transformed to butyric acid,
which provides between 70% and 90% of the energy required by colonocytes metabolism [22], thus sparing
about 30% of the energy requirement of piglets during the stressful period weaning represents [1] [22] [23].
Moreover, comparing results in the present study with other conducted by our research group, in which a low-
K. E. García et al.
7
Figure 2. Effects of dietary protein levels and probiotics on the
performance of newly weaned piglets. HPa: high-protein diet with
antibiotics; HP: high-protein diet without antibiotics; LPpb: low-
protein diet without antibiotics and with probiotics; ab: bars with
different letters show statistical differences; ADG1: average daily
gain in week 1; ADG2: average daily gain in week 2; ADG3: av-
erage daily gain in week 3; TADG: average daily gain during the
total experimental period; ADFI1: average daily feed intake in
week 1; ADFI2: average daily feed intake in week 2; ADFI3: av-
erage daily feed intake in week 3; TAFI: average daily feed intake
during the total experimental period; G:F1: gain: feed ratio in
week 1; G:F2: gain: feed ratio in week 2; G:F3: gain: feed ratio in
week 3; TG:F: total gain: feed ratio.
protein diet without probiotics was used (unpublished data), we can conclude that probiotics supplementation of
a low protein diet resulted in a higher proportion of SCFAs, mainly acetic acid, which could be beneficial to the
intestinal environment and probably reduce postweaning diarrhea in piglets.
K. E. García et al.
8
Furthermore, high concentration of SCFAs reduces gut pH, inhibiting the development of potentially patho-
genic bacteria [24]. The higher concentration of LA in the ileum with respect to that in the colon, may be the
result of rapid lactose fermentation in the small intestine, causing a reduction in lactose transit to the large bowel.
LA also serves as a bacterial control and promotes growth of beneficial bacteria, mainly SCFAs producers [1]
[2].
4.2. Postweaning Diarrhea
The high incidence and severity of diarrhea in piglets fed the HP diet suggest that high level of dietary protein
and the absence of antibiotic striggered postweaning diarrhea. This was also observed by others (5 - 9) and may
be explained as the fermentation of undigested protein by opportunistic bacteria, which results in an increased
production of BCFAs and AM, promoting the incidence and severity of diarrhea [25] [26].
Addition of antibiotics to high-protein diets controls bacterial growth, reduces overall fermentation, and sup-
presses BCFA and AM production [21] [22]. Reduction of protein levels and addition of probiotics in the diet
results in a beneficial fermentation pattern, increasing SCFAs and LA, and reducing BCFA and AM concentra-
tions. These changes may reduce pH values and in turn decreasing growth of opportunistic bacteria [1] [21].
SCFAs, mainly butyrate, increase glucagon-like peptide 2 (GLP2) secretion by L cells in the ileum. GLP2 sti-
mulates cellular proliferation and maturation, probably allowing a quick recovery of intestinal epithelium, re-
ducing the incidence and severity of postweaning diarrhea [27].
The transient under consumption normally reported during the first three days after weaning, probably reduc-
es the presence of undigested protein in the intestinal lumen, thereby suppressing bacterial fermentation [28].
Consequently, fermentation should not be a determining factor in the incidence of diarrhea during the first week
after weaning [28]. However, once piglets increase feed intake (3 - 5 days after weaning) the presence of fer-
mentable substrates (mainly protein) in the intestinal lumen may promote the growth of pathogenic bacteria, re-
sulting in greater production of potentially toxic metabolites, which can damage the intestinal mucosaproducing
diarrhea [1]. The severity of diarrhea observed during the second week after weaning confirmed this mechanism.
Once the animal’s digestive capacity increases, the severity of diarrhea decreases [29]; this was observed during
the third week after weaning.
The increase in SCFAs observed in piglets fed with the combination of a low-protein diet and probiotics
(LPpb diet), compared to those produced by piglets fed a high protein diet, may be the result of intestinal envi-
ronmental changes favoring the establishment of beneficial microbes and limiting the growth of pathogenic bac-
teria [22]. These changes may reduce the incidence and severity of postweaning diarrhea and improve piglet’s
growth. On the other hand, probiotics facilitate the characteristics of the intestinal environment by maintaining
an acidic pH, which inhibits the growth of potentially pathogenic bacteria that generally require an alkaline pHto
grow [30] [31]. Beneficial bacteria rapidly proliferate to become more abundant through competitive exclusion,
occupying binding sites on the intestinal mucosa that could otherwise be occupied by pathogenic bacteria [32].
Establishment of beneficial bacteria controls pathogenic bacteria proliferation through bacteriocin production,
which ruptures the bacterial cell wall through a mechanism similar to that of several antibiotics [30]-[33]. On the
other hand, probiotics have demonstrated to modulate the immune system, regulating the expression of pro-in-
flammatory cytokines, which contribute to the control of postweaning diarrhea [32].
4.3. Performance
Low-protein diet contents did not affect performance (ADG, ADFI, and G:F) of piglets (Figure 2). This result
was consistent with those of Le Bellego and Noblet [34], who found that diets with low crude protein, supple-
mented with essential amino acids, were effective to maximize nutrient intake by piglets. Furthermore, a de-
crease in dietary protein (from 20.4% to 16.9%) did not affect the ADFI or ADG of piglets. In the present study,
protein levels were slightly lower (16%), and no differences were observed. Bikker et al. [28] observed that an-
imals fed low-protein diets (15%) had higher ADFI and ADG than animals fed high protein diets. In a study by
Reynolds and O’Doherty [35] feeding a low protein diet to piglets between 0 and 28 days postweaning, resulted
in reduced growth rates, probably due to amino acids deficiency, mainly lysine. However, when appropriate
amino acids were provided together with a low-protein diet, animal growth was not altered. In the present study,
the low protein diet was supplemented with essential amino acids, including lysine, methionine, tryptophan,
threonine, and valine; we believe this is the reason why no changes in animal performance was observed. These
K. E. García et al.
9
results are consistent with those of Reynoso et al. [36], who found that protein content in the diet could be re-
duced as long as synthetic amino acids are included to meet the nutritional requirements of piglets. Similarly,
Hansen et al. [37] observed the same ADG in piglets fed low (17%) and high-protein (21%) diets supplemented
with amino acids.
5. Conclusion
In the present study, the use of antibiotics reduced microbial fermentation by altering the proliferation of intes-
tinal bacteria. When piglets were fed a high protein diet VFAs concentration was increased, mainly BCFAs.
Conversely, the use of a low-protein diet with probiotics promoted SCFAs production. Both high-protein diets
with antibiotics and low-protein diets with probiotics promoted healthy intestinal environment, reducing the in-
cidence and severity of diarrhea. Therefore, it can be concluded that feeding a low protein diet combined with
probiotics to newly-weaned piglets may have similar effects on incidence and severity of diarrhea and perfor-
mance, to those fed a high-protein diet with antibiotics. A combination of a low-protein diet and probiotics may
be an effective alternative to the use of antibiotics in starter diets.
Acknowledgements
This study was partially supported by CONACyT grant CB-2012-01000000000179898 and by the Programa In-
tergral de Fortalecimien to Institucional, of the Mexican Ministry of Public Education (SEP).
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... Fermentation of undigested dietary protein and endogenous proteins in the large intestines yield putative toxic metabolites that can impair epithelial integrity and promote enteric disorders such as PWD [27]. Incidence and severity of PWD may also be influenced by addition of probiotics to the diet, which may change the fermentation profile and thus promote gut health [28]. Furthermore, medium chain fatty acids (MCFAs) can neutralize bacterial metabolites in the small intestine [29]. ...
... Therefore, alternative approaches to control PWD due to pathogenic E. coli should be explored. Several alternative strategies, such as adapted nutritional strategies (feed consistency, lower crude protein, digestible fibers and other dietary fibers), prebiotics, probiotics, organic acids, MCFAs, specific IgA antibodies and oral vaccination have been explored [19][20][21][22][23][24][25][26][27][28][29]33,34,[37][38][39][40]]. ...
... Incidence and severity of PWD may also be influenced by addition of probiotics to the diet, which may change the fermentation profile and thus promote gut health [28]. Furthermore, medium chain fatty acids (MCFAs) can neutralize bacterial metabolites in the small intestine [29]. ...
... Therefore, E. coli F4 vaccination might be one of the alternative strategies once ZnO is banned in the EU by 2022. Besides vaccination, several other alternative strategies, such as adapted nutritional strategies (feed consistency, lower crude protein, digestible fibers and other dietary fibers), prebiotics, probiotics, organic acids, MCFAs, specific IgA antibodies and oral vaccination have been explored [19][20][21][22][23][24][25][26][27][28][29]33,34,[38][39][40][41]. ...
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Introduction: Post-weaning diarrhea (PWD) in pigs is a worldwide economically important disease, which is frequently controlled using antibiotics. However, emergence of antimicrobial resistance in Escherichia coli strains urges the need for alternative control measures, such as adapted feeding strategies, pre- and probiotics, organic acids, MCFAs or immunization. Methods: Different alternative control strategies such as active immunization of piglets against PWD with an E. coli F4 vaccine (Coliprotec® F4; Elanco) combined with high energy and protein diets, addition of nutraceuticals (medium chain fatty acids (MCFAs), organic acids and additional fibers) or supplementation of ZnO were evaluated for their efficacy against PWD due to F4 enterotoxigenic E. coli (F4-ETEC) under field conditions. Results: ZnO-supplemented piglets had a lower overall end weight and lower average daily weight gain, as compared to E. coli vaccinated piglets. The E. coli vaccinated group with normal energy and protein diet had the lowest clinical scores, whereas piglets fed a ZnO-supplemented diet had intermediate fecal clinical scores. All E. coli vaccinated groups had a low number of antibiotic treatments. In the nutraceutical group, clinical scores were much higher, indicating more severe clinical diarrhea, which needed additional antibiotic intervention. Conclusions: The present study demonstrated the efficacy of an oral live non-pathogenic E. coli F4 vaccine for active immunization of piglets against PWD due to F4-ETEC under field conditions. Different feeding strategies had no significant effect on the clinical outcome and performance parameters of E. coli vaccinated piglets.
... El uso de dietas bajas en proteína combinadas con Bacillus (B). subtilis y B. licheniformis en cerdos al destete, permitió cambiar el patrón fermentativo de la microbiota intestinal aumentando la concentración de ácido acético, propiónico y butírico, y reduciendo la concentración de amoniaco en el intestino delgado y ciego, disminuyendo la incidencia y severidad de diarreas, y mejorando los parámetros productivos comparados con los cerdos que recibieron dietas altas en proteína más antibiótico (76) . La adición de ácido benzoico y una mezcla de B. subtilis y B. licheniformis en lechones al destete mantuvo los parámetros productivos, redujo los conteos de coliformes y la liberación de amoniaco en aguas residuales comparados con las respuestas de cerdos al destete alimentados con dietas que contenían antibióticos (77) . ...
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En esta revisión se hace una retrospectiva de las actividades de investigación realizadas en nutrición porcina por los investigadores del INIFAP en los 35 años de existencia del Instituto. El producto principal de esta actividad, ha sido el sentar las bases para una mejor alimentación del hato reproductor y del cerdo para abasto, enfocándola a resolver los problemas particulares de la porcicultura mexicana, con respecto al uso de las materias primas utilizadas en la alimentación del cerdo; así como la evaluación y mejora de la calidad de la canal, el enriquecimiento de la carne con metabolitos que incrementan su vida de anaquel y mejoran sus propiedades organolépticas. Asimismo, se hace una reflexión sobre los retos que enfrentará la nutrición porcina en el presente siglo, proponiendo las áreas que se tendrán que investigar para garantizar la sustentabilidad del sector, así como las acciones que los investigadores del programa y el INIFAP tendrán que realizar para responder adecuadamente a los desafíos a los que se enfrenta la porcicultura mexicana.
... Weanling piglets are more at risk of retarded growth as well as an increase in both morbidity and mortality in pigs [1,4]. As the weaned piglets have limited digestive capacity, which triggers fermentation of undigested protein by opportunistic pathogens (mainly Escherichia coli, Salmonella) normally existing in gastrointestinal tract (GIT) which leads to production of branch chain fatty acids (BCFA) and ammonia nitrogen (NH3-N) [2,5]. The common practice of supplementing antibiotics in livestock for improved animal performance was condemned due to its adverse effects on animal as well as human, the ultimate consumer of the animal produce. ...
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A number of feed additives are marketed to assist in boosting the pigs' immune system, regulate gut microbiota, and reduce negative impacts of weaning and other environmental challenges. Some minerals (zinc and copper), dietary nucleotide, Direct-fed microbial (DFM) and yeast, some microorganisms (Lactobacillus) and plant extracts are, when administered in adequate amounts, confer good health benefits onto the host, are a category of feed additives that can be used to replenish the gut microbial population while recuperating the host immune system.
... Fermentation of undigested dietary protein and endogenous proteins in the large intestines yield putative toxic metabolites that can impair epithelial integrity and promote enteric disorders such as PWD [29]. Incidence and severity of PWD may also be influenced by addition of probiotics to the diet, which may change the fermentation profile and thus promote gut health [30]. Furthermore, Medium Chain Fatty Acids (MCFAs) can neutralize bacterial metabolites in the small intestine [31]. ...
... The low ADG observed in the first week after weaning can be explained by the stress of weaning, which provokes deleterious physiological and immunological changes and generalized poor development (Campbell et al., 2013). In the second post-weaning week, the piglets in the C+, Sc, and Sb diet groups had an ADG within the parameters usually reported for piglets of the same age group on the experimental farm (Escobar et al., 2015), while those in the C− diet group were below these parameters. Weight gain and faecal consistency indicated that the animals were in good health, in accordance with the post-weaning phase and the diet consumed. ...
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Emerging bacterial resistance to antibiotics increases the need for effective alternatives to control intestinal inflammation and thus gut disorders in piglets. This study evaluated the effects of including Saccharomyces cerevisiae (strain 1026) and Saccharomyces boulardii (CNCM I-1079) as antibiotic alternatives in the starter diets of pigs on the concentrations of ileal inflammatory markers (nuclear factor-KB (NF-kB)), interleukin-6 (IL-6), tumour necrosis factor alpha (TNF-a), and interleukin-12 subunit p40 (IL-12p40), and villus height. Forty piglets were assigned to four experimental diets, namely basal diet (C-), basal diet with antibiotics (C+), basal diet with S. cerevisiae (Sc), and basal diet with S. boulardii (Sb). At 7 and 14 days post weaning, five piglets per diet group were euthanized to quantify the inflammatory markers and to measure villus height. The C- group exhibited the highest concentration of inflammatory markers and the most atrophied villi. The Sc group had intermediate values for both variables. The C+ group had the lowest values for inflammatory markers and the highest villus height was similar to that of the Sb group, which showed low concentrations of inflammatory markers, although not so low as those of the C+ group. Both yeasts could be used as antibiotic alternatives to reduce the use of antibiotics in pig starter diets. However, S. boulardii CNCM I-1079 supplementation controls inflammation and preserves intestinal mucosa more effectively than S. cerevisiae strain 1026.
... The composition of the gastrointestinal microbiota is affected by dietary nutrition intake, for example, the three main nutritional components, proteins, carbohydrates, and fats [5]. Protein is the most essential component of tissues in animals and humans, and, unlike carbohydrates, high concentrations of dietary protein intakes could result in several deleterious metabolites in the gut [6]. Amounts of researches that aimed at figure out the effects of protein on infantile and model animals have showed that the concentration and sources of dietary proteins are potential causes of diarrhea in mammals [7]. ...
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Infantile diarrhea is a serious public health problem around worldwide and results in millions of deaths each year. The levels and sources of dietary protein are potential sources of diarrhea, but the relationship between the pathogenesis causes of infantile diarrhea and protein intake remains poorly understood. Many studies have indicated that the key to understanding the relationship between the protein in the diet and the postweaning diarrhea of piglets is to explore the influences of protein sources and levels on the mammalian digestion system. The current study was designed to control diarrhea control by choosing different protein levels in the diet and aimed at providing efficient regulatory measures for infantile diarrhea by controlling the protein levels in diets using a postweaning piglets model. To avoid influences from other protein sources, casein was used as the only protein source in this study. Fourteen piglets (7.98±0.14 kg, weaned at 28 d) were randomly allotted to two dietary treatments: a control group (Cont, containing 17% casein) and a high protein group (HP, containing 30% casein). The experiment lasted for two weeks and all animals were free to eat and drink water ad libitum. The diarrhea score (1=normal; 3=watery diarrhea) and growth performance were recorded daily. The results showed that the piglets in HP group had persistent diarrhea during the whole study, while no diarrhea was noticed in the control groups. Also, the feed intake and body weights were reduced in the HP groups compared with the other group (P
... Interestingly, an increase in ammonia concentration has a detrimental effect on the health of the GIT and a negative effect on the growth and differentiation of intestinal epithelial cells [49,50]. Additionally, BCFAs and ammonia are toxic metabolites for the intestinal mucosa and most likely trigger PWND and the poor performance in piglets [39, 51,52]. More importantly, the upregulated expression of ammonia may induce a disorder of the intestinal microbial balance during weaning [20]. ...
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Piglets experience severe growth challenges and diarrhea after weaning due to nutritional, social, psychological, environmental, and physiological changes. Among these changes, the nutritional factor plays a key role in postweaning health. Dietary protein, fibre, starch, and electrolyte levels are highly associated with postweaning nutrition diarrhea (PWND). In this review, we mainly discuss the high protein, fibre, resistant starch, and electrolyte imbalance in diets that induce PWND, with a focus on potential mechanisms in weaned piglets.
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This study was conducted to determine the effects of low-protein diets supplemented with crystalline amino acids (AA) on performance and intestinal development in barrows (n=32) over 2 weeks after weaning at 18±1 day. Four maize-soybean meal based diets providing 0.93 g standardized ileal digestible lysine/MJ ME were prepared. The treatments were a control diet containing 23.1% CP (crude protein) and three low-protein diets (21.2, 18.9, and 17.2% CP, respectively), which were supplemented with crystalline AA to achieve an ideal AA pattern. Piglets were raised individually and had free access to feed and water. Average daily gain (ADG) and average daily feed intake (ADFI) were measured weekly. Severity of diarrhea was monitored twice per day. Blood from all piglets was taken for determining serum urea nitrogen on d 0, 7, and 14 and serum free AA concentration on d 14. At the end of the experiment, all piglets were killed to measure morphology of the small intestine and disaccharidase activities. Reducing CP level did not affect ADFI (P>0.10) but resulted in poorer (linear, P
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The objective of this experiment was to investigate the effect of non-starch polysaccharide (NSP) solubility and inclusion rate on gut health and development, performance and risk of post weaning enteric disorders (PWED) using NSP sources known not to affect digesta viscosity. The experiment consisted of a 2×3 factorial combination of NSP type (sNSP vs insoluble NSP (iNSP); inulin vs purified cellulose) and inclusion level (50 vs 100 vs 150 g/kg). Piglets were weaned at 28 days (day 0) and fed ad libitum until slaughter on day 14. There were no incidences of PWED. NSP solubility had little effect on performance, but sNSP diets resulted in lower caecal and colonic pH and higher colonic Lactobacilli to coliform ratio compared to iNSP diets. Increasing NSP levels significantly reduced caecal and colonic pH but decreased empty body weight percentage (P=0.067). These results suggest that gut health may benefit most from diets containing appropriate sources of predominantly sNSP rather than iNSP. The optimal level of such sNSP will likely depend upon the level of risk of PWED in order to balance the trade off between maximising performance and improving gut health. Effects of NSP feeding under a higher infectious pressure remains to be assessed.