Piglet Feed Contaminated with Aatoxin B1 and Supplemented with an Anti-Mycotoxin Blend: Impact on Health and Zootechnical Performance

The present study aimed to determine whether the inclusion of a mixture based on Saccharomyces cerevisiae lysate, zeolite, silicon dioxide, propylene glycol, Carduus marianus extract, soy lecithin, and carbonate in the piglet diet would be able to minimize the negative effects caused by feed with aatoxin; a study focusing on animal health and performance. Seventy-two entire male piglets (7.35 ± 1.17 kg) weaned with an average of 26 days were used, allotted into four groups, with six repetitions (pens) per group and three piglets/pen. The treatments were as follows: NoAa-NoAntiMyc - negative control (without aatoxin); Aa-NoAntiMyc - positive control (500 ppb of aatoxin); NoAa-AntiMyc - 1000 mg/kg of anti-mycotoxin mixture; Aa-AntiMyc - 500 ppb aatoxin + 1000 mg/kg anti-mycotoxin. We evaluated the performance (feed intake - FI, weight gain - WG and feed:gain ratio- FGR) andblood variables (10 days between samples). On day 32, six pigs from each group (a total of 24 pigs) were slaughtered to collect fragments of the liver, intestine, and spleen for analysis of oxidants/antioxidants and histology. It was observed that piglets in the positive control group (Aa-NoAntiMyc) had lower body weight and weight gain when compared to the other groups during the experimental period. Also, piglets from Aa-NoAntiMyc consumed less feed between days 1 to 20 and 1 to 30 compared to NoAa-NoAntiMyc. A considerable increase in liver enzymes aspartate aminotransferase and alanine aminotransferase in the piglets' serum of Aa-NoAntiMyc compared to other treatments, which indicates that the anti-aatoxin blend worked. Fewer neutrophils and an increase in monocyte were observed in the piglets of Aa-NoAntiMyc compared to NoAa-NoAntiMyc; changes that were not seen in the animals of Aa-AntiMyc. We did not see any change in the oxidant and antioxidant status in the piglet serum during the experimental period; different from what happened in Aa-NoAntiMyc piglets in the liver (higher glutathione reductase activity and levels of reactive oxygen species - ROS), in the spleen (higher levels of ROS) and the intestine/jejunum (reduction of nitrate/nitrite levels - NOx). Intestinal morphometry revealed that piglets from Aa-NoAntiMyc had higher villus height than the other groups, while the folded size was smaller in this group. The crypt depth was greater in the intestine of piglets in both treatments that consumed aatoxin. In general, it is concluded that the consumption of aatoxin B1 by piglets has negative impacts on the health and, consequently, the animals' performance; however, supplementation of the contaminated feed with an anti-mycotoxin blend was able to protect the piglets, minimizing the negative problems caused by the mycotoxin.

nursery and growing phase, morbidity reached 97%, among 68 of the 70 animals; however, the younger animals were more susceptible to a atoxicosis outbreaks (Schoenau, 1994).
Foods contaminated with a atoxin directly reach productivity, which has forced the search for strategies to prevent or reduce this intoxication in animals (Ramos, 1996). One of the productive sector alternatives is prophylactically protecting animals by adding adsorbents to the feed (Huwig, 2001). These adsorbents are also known as mycotoxin binders and sequestering as agents that act directly on the intestinal tract, decreasing the absorption of these toxins and promoting their excretion through urine or feces (Haque, 2020). Considering that the use of adsorbents is an effective way and one of the main methods used to prevent possible pathologies arising from the ingestion of mycotoxins in piglets (Bretas, 2018), the present work brought as a strategy the use of a blend that among its ingredients has the C. marianum, an extract rich in silymarin, with the hypothesis that it would minimize adverse effects caused by a atoxin, due to acting as a hepatoprotective (Magliulo, 1979;Lorenz, 1984;Valenzuela, 1985;Yao-Cheng, 1991;Conti, 1992;Muriel, 1992;Haková, 1993;Wu, 1993). Another ingredient in the blend was Saccharomyces cerevisiae lysate, which has high levels of complex B vitamin and can stimulate the immune system, which has already resulted in better feed:gain ratio and weight gain (Simon, 2001); and soy lecithin with its antioxidant action (Marconcin, 2009); among the other strategic ingredients.
We hypothesize that the blend can neutralize the adverse effects of experimental a atoxicosis, directly or indirectly, avoiding production losses in cases of contamination of piglets' food by a atoxin. Thus, the objective of this research was to determine whether the inclusion of a mix based on S. cerevisiae lysate, zeolite, silicon dioxide, propylene glycol, C. marianus extract, soy lecithin, and carbonate in the piglet diet would be able to minimize the adverse effects caused by the daily intake of a atoxin; this study focused on pig performance and health.

Anti-mycotoxin commercial product
We used commercial products based on S. cerevisiae lysate, zeolite, silicon dioxide, propylene glycol, C. marianus extract, soy lecithin, and calcium carbonate.

A atoxin production and analysis
A atoxins were produced by the ATCC 13608 strain of Aspergillus avus during fermentation of converted rice, and the following protocol was used. Erlenmeyer asks of 500 mL volume were used to receive 100 g of rice. At least 2 h before the sterilization of 40 distilled water was added to a ask and mixed with rice. The sterilization was performed at 121 ºC for 30 minutes, and then the asks were left to lose temperature before inoculation. The rice was inoculated with 2 mL of 108 spore mL − 1 of spore suspension of A. avus. The incubation was carried out for 21 days at a controlled temperature (25 ºC) and constant stirring of asks. After incubation, the fermented material was dried in an oven at 50 ºC and ground. The concentration of a atoxin in the inoculum was determined in advance in order to calculate and determined the amount added in the diets in order to obtain a 500 ppb contamination, a dose already described in the literature for impairing pig performance (Schell et al., 1993).
Samples of feed and inoculum were ground to < 0.85 mm material, and 1 g of the ground material was transferred to a test tube of 50 mL. It was added 10 mL of ultrapure water and 10 mL of acetonitrile/acetic acid (CH3CN: CH3COOH) [99.5:0.5, v/v], and the test tube was placed in a mechanic shaker for 10 min. A mixture of 4 g of MgSO4 and 1 g of NaCl was, added and the tube was vigorously shaken for 10 s. The solution was centrifuged for 15 min at 5.000 x g, at, 25 •C and 2.5 mL of supernatant was transferred to capped glass test tube where 2.5 mL of hexane was added. The solution was shaken for 2 h and then centrifuged at 1,000 x g at 20 ºC for 1 min. The lower phase (acetonitrile) 1 mL was withdrawn and dried with a nitrogen (N2) stream at 40 ºC. The reconstitution was performed with 75 µL of methanol in an ultrasonic bath for 10 s and 10 s in a test tube mixer after adding 75 µL of ultrapure water. After centrifugation for 10 min at 14,000 x g, 60 µL was withdrawn and transferred to a vial where 140 µL of ultrapure water was added. Ten microliters were injected into a chromatographic system. Detection and quanti cation of a atoxins were performed with high-performance liquid chromatography coupled with a mass spectrometer (LC/MS/MS). Chromatographic separation was carried out using Acqulty UPLC System (Waters, Milford, Massachusetts, US) equipped with 100 × 2.1 mm, 1.7 µm Acquity UPLC BEH C18 column (Waters, Milford, Massachusetts, US). The column was maintained at 40 •C, and the injection volume was 10 µL. The mobile phase consisted of 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B). The acetonitrile (B) concentration was raised gradually from 10-90% within 12 min, brought back to the initial conditions at 0.1 min, and allowed to stabilize for 3 min. The mobile phase was delivered at a ow rate of 0.4 mL/min. The LC system was coupled with Xevo TQS tandem mass spectrometer (Waters, Milford, Massachusetts, US), equipped with a turbo-ion electrospray (ESI) ion source. The mass spectrometer was operated in scheduled multiple reaction monitoring (MRM) in positive mode. The electrospray ionization and MS/MS conditions as shown in Supplementary Material 1.

Animals and experimental design and Nutritional analysis
The experiment was carried out in the experimental pig facilities at the Experimental Farm of the State University of Santa Catarina (FECEO), located in Guatambu, SC, Brazil, over 40 days. According to the nutritional requirements of pigs, the diet (mash) was based on corn, soybean meal, and commercial basemix to meet the nutritional requirements of pigs. Previous the corn grain was cleaning by rotary sieve machine (5 mm of aperture of sieves) to remove broken grains, sand and other small impurities.
We used 72 entire male piglets (7.35 ± 1.17 kg) weaned with an average of 26 days, allotted into four groups with six replicates (six pens with 3 piglets). The experiment was conducted in a nursery facility tted with a plastic oor, linear feeder (15cm/pigs) nipple drinker (minimum ow rate 1/L/min). The facilities' temperature was maintained with a convective automated heating system (5,400 W heaters), activate at 28°C and deactivate at 29°C, additional heaters (1,200 W) were positioned in the corner's facilites (with the same set) as additional measure to meet the adequate temperature the temperature program was reduced 1ºC per week ( Supplementary Fig. 1). Dry bulb temperature was recorded using dataloggers (Probe type 18b20) positioned in the geometric center of the installation with 1 h sampling intervals ( Supplementary Fig. 1).
The treatments were as follows: NoA a, negative control (without a atoxin); A a, positive control (500 ppb of a atoxin); Anti-Myc, 1000 mg/kg of the anti-mycotoxin blend; and A a + Anti-Myc, 500 ppb of a atoxin + 1000 mg/kg of A a + Anti-Myc.
The diets (mash form) from each phase and treatment were ground (mesh 1 mm) and analyzed dry matter (DM,

Zootechnical performance
The zootechnical performance was evaluated at the end of the steps I (0-10 days), II (11-20 d), III (21-30 d) and IV (31-40 d) of the experimental periods. Pig weight were recorded an electronic scale (model DIGI-TRON UL-5 ± 50g) and feed intake recorded an electronic scale (Toledo ± 5g). Were calculated the daily feed intake, daily weight gain and feed:gain ratio calculated per pen

Sample collection
Blood samples were collected in vacutainer tubes on days 0, 10, 20, 30, and 40 of the experimental periods in tubes containing anticoagulants by trained staff. First, complete blood counts were performed according to the methodology described below. A 0.5 mL aliquot of blood was removed to analyze catalase (CAT) and superoxide dismutase (SOD) activity and stored frozen. Subsequently, blood was centrifuged at 8,000 rpm for 5 minutes, thereby obtaining serum allocated in a microtube and maintained frozen (-20°C) until biochemical analysis.
On day 32 of the experiment, six pigs from each group were slaughtered in a specialized slaughterhouse, according to the inspection system's current legislation. Fragments of the liver, intestine, and spleen were collected, and samples were preserved in 10% formaldehyde. A liver fragment was homogenized in saline, centrifuged, and the supernatants were removed. These were packed in microtubes and frozen for further analysis of oxidants/antioxidants.

Hemogram
According to the manufacturer's recommendations, the hemoglobin, total leukocyte, and erythrocyte contents were determined using a commercial kit. In the sampling, blood smears were made and stained with commercial dye (Rapid Panotype) to perform differential leukocyte counts under a light microscope with a 1000 x magni cation, as described by Lucas and Jamroz (1961). Hematocrit was measured using microcapillary tubes, centrifuged at 14000 x g for 5 min.

Serum biochemistry
Serum levels of total proteins, albumin, cholesterol, triglycerides, alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were measured using semi-automatic BioPlus equipment (Bio-2000) and speci c commercial kits. Serum globulin levels were calculated as the difference between serum levels of total proteins and albumin.
GST activity was measured according to Mannervik and Guthenberg (1981), with modi cations. Brie y, GST activity was measured as the rate of formation of dinitrophenyl-S-glutathione at 340 nm in a medium containing 50 mM potassium phosphate at pH 6.5, 1 mM GSH, 1 mM 1-chloro-2, 4-dinitrobenzene (CDNB) as substrate and tissue supernatants (approximately 0.045 mg protein). The results were expressed as U GST/mg protein. The activity of the SOD was measured using the method of Marklund and Marklund (1974), and the results were expressed as nmol SOD/mg of protein. CAT activity was measured using ultraviolet spectrometry, according to the method described by Aebi (1984), and the results were expressed as nmol CAT/mg of protein.
The levels of reactive oxygen species (ROS) in plasma were analyzed by the method described by Halliwell and Gutteridge (2007). The plasma (10 µL) was incubated with 12 µL of dichloro uorescein (DFC) per mL at 37 ° C for 1 h in the dark. Fluorescence was determined using 488 nm for excitation and 520 nm for emission. The results were

Organ weight and histopathology
Spleen and liver were weighed during the slaughter process. Then, fragments of the liver, intestine, and spleen were preserved in a formaldehyde solution (10%). Tissue fragments were processed and placed in para n blocks. Then sections were made and stained with hematoxylin-eosin (HE).

Statistical analysis
The experimental design of this study was one factorial 2 × 2 [feed with and without a atoxin (A a and NoA a) and with or without anti-mycotoxin (Anti-Myc and NoAnti-Myc)]. All data were analyzed using the MIXED procedure of SAS (SAS Inst. Inc., Cary, NC, USA; version 9.4), with Satterthwaite approximation to determine the denominator degrees of freedom for the test of xed effects. The data of feed intake, weight gain, feed:gain ratiowere tested for xed effects of a atoxin, silymarin, and the interaction, and as a random effect was included pen (a atoxin × silymarin). The data of antioxidant response in liver, spleen, and intestines variables were tested for xed effects of a atoxin, silymarin,n, and the interaction. Ass random effects were included pen (a atoxin × anti-mycotoxin) and pigs (pen). All other data were analyzed as repeated measures (body weight and blood variables) and were included as xed effects a atoxin, antimycotoxin, day and all possible interactions, and as random effects were included pen (a atoxin × anti-mycotoxin) and pig (pen). The covariance structures were selected according to the lowest Akaike information criterion. Means were separated using PDIFF, and all results were reported as LSMEANS followed by SEM. Signi cance was de ned when P ≤ 0.05 and tendency when P > 0.05 and ≤ 0.10.

Performance
The body weight and weight gain of pigs that consumed feed contaminated with a atoxin were lower than the other treatments ( Table 2). DWG had an a atoxin effect (days 1 to 10; 1 to 20, 1 to 30) and had an interaction between a atoxin versus anti-mycotoxin (days 1 to 10; 1 to 30 and 21 to 30), i.e., piglets of the A a-NoAntiMyc groups had lower DWG. Daily feed intake was lower in the A a-No Anti Myc group in periods 1 to 20 and 1 to 30, which characterizes an a atoxin effect (Table 2). There was no effect of the treatment and additive on feed conversion ratio.

Serum biochemistry
There was day versus AntiMyc interaction for total protein and globulin levels on days 20 and 40 of the experiment. Both variables had a higher concentration in the piglet serum that consumed the tested anti-mycotoxin blend (Table 3).
Albumin levels were affected by the consumption of feed contaminated with a atoxin, i.e., albumin levels in the serum of these piglets were higher (Table 3). Triglycerides and cholesterol did not differ between treatments, with no effect of a atoxin and anti-mycotoxin blend.
ALT and AST activity affected a atoxin and the anti-mycotoxin blend; there was an interaction of these variables with day (Table 3). We emphasize that a atoxin consumption increased the aforementioned liver enzymes' activity, but when the anti-mycotoxin blend was added to the piglet's diet, the adverse effects on AST and ALT were avoided (Table 3; Figure 1).

Hemogram
There was no effect of a atoxin consumption and anti-mycotoxin blend on erythrogram (total erythrocytes, hemoglobin, and hematocrit) ( Table 4). There was also no effect of a atoxin intake and anti-mycotoxin blend on total leukocytes, lymphocytes, and eosinophils during the experiment period; different from what happened with the number of neutrophils and monocytes ( Table 4).
The consumption of a atoxin by piglets caused a lower neutrophil count during the experimental period. There was a tendency toward a lower neutrophil count when the pigs consumed the anti-mycotoxin blend ( Table 4). The consumption of the anti-mycotoxin blend in uenced the monocyte count, which was lower (Table 4); in addition, there was an interaction between a atoxin and anti-mycotoxin characterized by a higher monocyte count in piglets that ingested a atoxin (Figure 2).

Plasma, blood, and tissue antioxidant responses
In the serum, antioxidant enzymes (GST, SOD, and CAT) and oxidizing biomarkers (NOx, ROS, and TBARS) did not differ between treatments, without the effect of a atoxin and the consumption of the anti-mycotoxin blend ( Table 5).
In the liver, there was an interaction between a atoxin versus anti-mycotoxin for GST activity and ROS levels ( Table 6); both variables had higher values (A a-NoAntiMyc) when compared to other treatments. There was an interaction between a atoxin versus anti-mycotoxin for ROS levels; that is, the piglets that consumed a atoxin had higher ROS levels than other treatments (Table 6).
In the intestine, we veri ed the effect of the consumption of a atoxin and the consumption of anti-mycotoxin for NOx levels; that is, piglets that consumed a atoxin had higher NOx levels when compared to other treatments (Table 6). A atoxin led to a reduction in intestinal NOx levels, differently from the consumption of anti-mycotoxin blend, which increased NOx levels in the intestine (Table 6).

Histopathology
No intestinal, hepatic, and spleen lesions were observed in any treatment. There was an interaction between a atoxin and anti-mycotoxin blend for fold size, villus height, and crypt depth (Table 7). A atoxin consumption decreased the fold-size but increased villus height and crypt depth. The consumption of the anti-mycotoxin blend minimized these changes regarding the fold and villus (Table 7).

Discussion
The consumption of diets with a atoxin by pigs impaired weight gain, which was expected and made it possible to validate this study. According to Dilkin (2002), the ingestion of moderate or low doses of mycotoxins causes chronic mycotoxicosis interferes with productive performance, reducing the rate of feed conversion, weight gain, and reproductive e ciency. In swine herds that consumed different concentrations of a atoxin, changes have already been described, such as lower feed consumption and gain and weight, in addition to higher mortality in higher concentrations (Cook, 1989). Wang (2020), in a recent survey of piglets, weaned and fed with diets contaminated with different levels of a atoxin (0, 250, and 500 ppb), observed that as the dose of mycotoxin increased, the rate of performance of these pigs decreased linearly, reaching a 25% reduction in GMD when administered with a diet containing 500 ppb of a atoxin, which is already known (Dilkin, 2002;Cook, 1985).
The anti-mycotoxin blend used in the present study minimized the adverse effects on weight gain caused by a atoxin.
The addition of adsorbents in animal feed to protect animals is a common practice (Huwig, 2001); however, blends have been a multifunctional alternative, that is, in addition to minimizing the effects of mycotoxins, it also has properties biological. Weaver (2013) evaluated the performance and health of pigs challenged by mycotoxins and fed simultaneously by three different types of adsorbents. It was found that the addition of adsorbents improved the health of the pigs and also favored the growth rate. This can be explained by the fact that a atoxin, when absorbed by the gastrointestinal tract, can cause depression, diarrhea, and inappetence, reducing food conversion and consequently weight gain (Dilkin, 2004).
However, when an adsorbent blend is used in the feed of pigs that are eating contaminated food, the adsorbent acts directly on the pigs's body, xing itself on the mycotoxin and eliminating it in the urine and feces, preventing its absorption (Bretas, 2018) and consequently not compromising and impairing the pig's performance. In this speci c case, it is possible to highlight the contribution of silymarin present in the composition of the blend used, as it has an anti-hepatotoxic action, stabilizes the membrane, protects and prevents lipid peroxidation (Bindoli, 1997, Magliulo, 1979, Yao-Cheng, 1991, Lastra, 1992, Muriel, 1992. Considerable increase in liver enzymes (AST and ALT) in the serum of piglets that consumed a atoxin was observed, which did not occur in the pigs' serum when they consumed the anti-mycotoxin blend; therefore, how directly or indirectly did it have a hepatoprotective effect, capable of preventing cellular damage to the liver. According to the literature, the consumption of a atoxin by animals generates liver problems (De La Salud 1993;Shen, 1995;Lopez, 2002;Kamdem, 2006;Yarru, 2009). When the animal is intoxicated by ingesting products contaminated by a atoxin, it is highly susceptible to several liver problems since it is the main target (Dilkin, 2004).
Among the most frequent problems that may occur are injuries de ned by fatty degenerations, the proliferation of bile ducts, biochemical changes, and blood clotting (Borsa, 1998). According to Olinda (2016), when a signi cant increase in the enzymes ALT and AST is observed, it suggests possible liver problems (hepatocellular lesions) of acute, hyperacute, or chronic level, since these are synthesized at the mitochondrial level. It is essential to investigate and understand how the anti-mycotoxin blend was able to protect the liver, which may be related to the presence of C. marianum, an extract rich in silymarin, a component known for its hepatoprotective effect (Ferreira, 2011).
However, another blend similar to the one used here, containing S. cerevisiae lysate, was tested in laying hens challenged with FB1 and DON, with a protective effect on the liver (Dazuk, 2021, in press). Despite cellular damage caused by a atoxin, the liver's functionality was not affected by interfering with production. However, the synthesis of albumin was increased in the serum of piglets that consumed a atoxin. Other researchers have already described that as the concentration of the toxic substance (mycotoxins) in the food offered increased, albumin levels increased proportionally (Rotter, 1994).
Lower neutrophil counts were observed in the readers of the A a500Anti0 group, a change not observed in the pigs of the A a500Anti1000 group. As mycotoxin acts as an immunosuppressant (Sharma 1993), we believe that the blend was also able to avoid this hematological alteration resulting from the consumption of a atoxin. However, the number of monocytes was lower in piglets in the A a500Anti1000 group. Today it is known that humoral immunity is affected by several mycotoxins (Oswald, 1998;Bonde, 2000). In an evaluation of sows exposed to mycotoxins, macrophage and neutrophil activities were inhibited (Silvotti, 1997). An experiment with piglets in daycare exposed to food contaminated with a atoxin demonstrated a lower activity of messenger RNA in pro-in ammatory and anti-in ammatory cytokines (Oswald, 2005), which also suggests our results.
GTS activity and ROS levels in the liver were elevated in the group of piglets exposed to mycotoxin (A a500Anti0), similar to what happened in the spleen to concentrate ROS. In pig farming, oxidative stress is a signi cant problem in newly weaned piglets, as these piglets are in a more delicate period and are more susceptible to this problem (Amazan, 2012). According to Shi-Bin (2007), this stress in piglets compromises their entire development since it will reduce their digestibility of nutrients and consequently compromising their growth. When this imbalance occurs, the increase in ROS occurs, increasing the animal's susceptibility to triggering infectious diseases since this alteration compromises its immune system when at signi cantly exacerbated levels (Bergman, 2011). It is known that in pigs, one of the factors that corroborate for this oxidant-antioxidant imbalance to occur is the animals' diet, so it is essential to know the quality of the food available (Falowo, 2014); because ingredients and consequently food contaminated by mycotoxins cause oxidative stress for example (Hou, 2013), as well as compromising biological functions.

Conclusion
A atoxicosis in pigs' impaired growth and concludes that the tested anti-mycotoxin blend has a protective effect on the health of the pigs and, consequently, minimizes the a atoxin effects in contaminated diets. The results suggest that the blend acts indirectly, differently from conventional adsorbents; that is, the blend, when consumed by animals, has biological properties capable of protecting the liver, the metabolic organ, mainly affected by intoxications a atoxin.

Declarations Data availability
All data and materials used in the experiment are available and are ready to be provided if needed.

Consent to participate
All names in the author list have been involved in various stages of experimentation or writing.

Consent for publication
All authors agree to submit the paper for publication in the Tropical Animal Health and Production.    2 In a factorial design (2 × 2) was include or not a atoxin (NoA a and A a for 0 or 500 mg of a atoxin/kg of concentrate, respectively) and also included or not anti-mycotoxin blend (NoAntiMyc and AntiMyc for 0 or 1000 mg of anti-mycotoxin blend/kg of concentrate, respectively). 3 A a, a atoxin; AntiMyc, anti-mycotoxin blend. a-b Differs (P ≤ 0.05) between treatments (lines).  2 In a factorial design (2 × 2) was include or not a atoxin (NoA a and A a for 0 or 500 mg of a atoxin/kg of concentrate, respectively) and also included or not anti-mycotoxin blend (NoAntiMyc and AntiMyc for 0 or 1000 mg of anti-mycotoxin blend/kg of concentrate, respectively). 3 A a, a atoxin; AntiMyc, anti-mycotoxin blend.
a-b Differs (P ≤ 0.05) between treatments (lines).  2 In a factorial design (2 × 2) was include or not a atoxin (NoA a and A a for 0 or 500 mg of a atoxin/kg of concentrate, respectively) and also included or not anti-mycotoxin blend (NoAntiMyc and AntiMyc for 0 or 1000 mg of anti-mycotoxin blend/kg of concentrate, respectively). 3 A a, a atoxin; AntiMyc, anti-mycotoxin blend.
a-b Differs (P ≤ 0.05) between treatments (lines).  2 In a factorial design (2 × 2) was include or not a atoxin (NoA a and A a for 0 or 500 mg of a atoxin/kg of concentrate, respectively) and also included or not anti-mycotoxin blend (NoAntiMyc and AntiMyc for 0 or 1000 mg of anti-mycotoxin blend/kg of concentrate, respectively). 3 A a, a atoxin; AntiMyc, anti-mycotoxin blend.
a-b Differs (P ≤ 0.05) between treatments (lines). Table 7. Intestinal morphometry of piglets exposed to a atoxin and supplemented with anti-mycotoxin blend.

Figure 1
Serum concentration of alanine aminotransferase (ALT) and aspartate aminotransferase (AST of piglets fed with diets containing a atoxins (A a) and anti-mycotoxin blend (Anti-Myc). In a factorial design (2 × 2) was include or not a atoxin (NoA a and A a for 0 or 500 mg of a atoxin/kg of concentrate, respectively) and also included or not antimycotoxin blend (NoAntiMyc and AntiMyc for 0 or 1000 mg of anti-mycotoxin blend/kg of concentrate, respectively). a-cDiffers (P ≤ 0.05) between treatments (lines). Vertical bars represent the SEM.

Figure 2
Number of monocytes of piglets fed with diets containing a atoxins (A a) and anti-mycotoxin blend (Anti-Myc). In a factorial design (2 × 2) was include or not a atoxin (NoA a and A a for 0 or 500 mg of a atoxin/kg of concentrate, respectively) and also included or not anti-mycotoxin blend (NoAntiMyc and AntiMyc for 0 or 1000 mg of anti-mycotoxin blend/kg of concentrate, respectively). a-cDiffers (P ≤ 0.05) between treatments (lines). Vertical bars represent the SEM.

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