Assessment of the Impact of Humic Acids on Intestinal Microbiota, Gut Integrity, Ileum Morphometry, and Cellular Immunity of Turkey Poults Fed an Aflatoxin B1-Contaminated Diet.

A recent study published data on the growth performance, relative weights of the organs of the gastrointestinal tract, liver histology, serum biochemistry, and hematological parameters for turkey poults fed an experimental diet contaminated with aflatoxin B1 (AFB1) and humic acids (HA) extracted from vermicompost. The negative effects of AFB1 (250 ng AFB1/g of feed) were significantly reduced by HA supplementation (0.25% w/w), suggesting that HA might be utilized to ameliorate the negative impact of AFB1 from contaminated diets. The present study shows the results of the remaining variables, as an extension of a previously published work which aimed to evaluate the impact of HA on the intestinal microbiota, gut integrity, ileum morphometry, and cellular immunity of turkey poults fed an AFB1-contaminated diet. For this objective, five equal groups of 1-day-old female Nicholas-700 turkey poults were randomly assigned to the following treatments: negative control (basal diet), positive control (basal diet + 250 ng AFB1/g), HA (basal diet + 0.25% HA), HA + AFB1 (basal diet + 0.25% HA + 250 ng AFB1/g), and Zeolite (basal diet + 0.25% zeolite + 250 ng AFB1/g). In the experiment, seven replicates of ten poults each were used per treatment (n = 70). In general, HA supplementation with or without the presence of AFB1 showed a significant increase (p < 0.05) in the number of beneficial butyric acid producers, ileum villi height, and ileum total area, and a significant reduction in serum levels of fluorescein isothiocyanate-dextran (FITC-d), a marker of intestinal integrity. In contrast, poults fed with AFB1 showed a significant increase in Proteobacteria and lower numbers of beneficial bacteria, clearly suggesting gut dysbacteriosis. Moreover, poults supplemented with AFB1 displayed the lowest morphometric parameters and the highest intestinal permeability. Furthermore, poults in the negative and positive control treatments had the lowest cutaneous basophil hypersensitivity response. These findings suggest that HA supplementation enhanced intestinal integrity (shape and permeability), cellular immune response, and healthier gut microbiota composition, even in the presence of dietary exposure to AFB1. These results complement those of the previously published study, suggesting that HA may be a viable dietary intervention to improve gut health and immunity in turkey poults during aflatoxicosis.

Evaluation of the Antimicrobial Activity of a Formulation Containing Ascorbic Acid and Eudragit FS 30D Microparticles for the Controlled Release of a Curcumin-Boric Acid Solid Dispersion in Turkey Poults Infected with Salmonella enteritidis: A Therapeutic Model.

The selection of components within a formulation or for treatment must stop being arbitrary and must be focused on scientific evidence that supports the inclusion of each one. Therefore, the objective of the present study was to obtain a formulation based on ascorbic acid (AA) and Eudragit FS 30D microparticles containing curcumin-boric acid (CUR-BA) considering interaction studies between the active components carried out via Fourier transform infrared spectrometry (FTIR) and differential scanning calorimetry (DSC) to minimize antagonistic effects, and comprehensively and effectively treat turkey poults infected with Salmonella enteritidis (S. enteritidis). The DSC and FTIR studies clearly demonstrated the interactions between AA, BA, and CUR. Consequently, the combination of AA with CUR and/or BA should be avoided, but not CUR and BA. Furthermore, the Eudragit FS 30D microparticles containing CUR-BA (SD CUR-BA MP) showed a limited release of CUR-BA in an acidic medium, but they were released at a pH 6.8-7.0, which reduced the interactions between CUR-BA and AA. Finally, in the S. enteritidis infection model, turkey poults treated with the combination of AA and SD CUR-BA MP presented lower counts of S. enteritidis in cecal tonsils after 10 days of treatment. These results pointed out that the use of an adequate combination of AA and CUR-BA as an integral treatment of S. enteritidis infections could be a viable option to replace the indiscriminate use of antibiotics.

Curcumin reduces enteric isoprostane 8-iso-PGF2α and prostaglandin GF2α in specific pathogen-free Leghorn chickens challenged with Eimeria maxima.

The purpose of this pilot study was to evaluate and determine the concentration of prostaglandin GF2α (PGF2α) and isoprostane 8-iso-PGF2α in plasma and intestine of specific pathogen-free (SPF) Leghorn chickens challenged with Eimeria maxima, with or without dietary supplementation of curcumin using solid-phase microextraction and ultra-performance liquid chromatography/tandem mass spectrometry. Eighty 1-day-old male SPF chickens were randomly allocated to one of four groups with four replicates (n = 5 chickens/replicate). Groups consisted of: (1) Control (no challenge), (2) Curcumin (no challenge), (3) Eimeria maxima (challenge), and (4) Eimeria maxima (challenge) + curcumin. At day 28 of age, all chickens in the challenge groups were orally gavaged with 40,000 sporulated E. maxima oocysts. No significant differences (P > 0.05) were observed in the groups regardless of the treatment or challenge with E. maxima. Enteric levels of both isoprostane 8-iso-PGF2α and PGF2α at 7 days and 9 days post-challenge were significantly increased (P < 0.01) compared to the non-challenge control chickens. Interestingly, the enteric levels of both isoprostane 8-iso-PGF2α and PGF2α at 7 days post-challenge were significantly reduced in chickens fed curcumin, compared to control chickens challenge with E. maxima. At 9 days post-challenge, only levels of isoprostane 8-iso-PGF2α in the enteric samples were significantly reduced in chickens challenged with E. maxima supplemented with curcumin, compared with E. maxima challenge chickens. No differences of isoprostane 8-iso-PGF2α or PGF2α were observed in plasma at both days of evaluation. Similarly, no significant differences were observed between the challenge control or chickens challenge with E. maxima and supplemented with curcumin at both times of evaluation. The results of this pilot study suggests that the antioxidant anti-inflammatory properties of curcumin reduced the oxidative damage and subsequent intestinal mucosal over-production of lipid oxidation products. Further studies to confirm and extend these results in broiler chickens are required.

Evaluation of curcumin and copper acetate against Salmonella Typhimurium infection, intestinal permeability, and cecal microbiota composition in broiler chickens.

BACKGROUND: Interest in the use of natural feed additives as an alternative to antimicrobials in the poultry industry has increased in recent years because of the risk of bacterial resistance. One of the most studied groups are polyphenolic compounds, given their advantages over other types of additives and their easy potentiation of effects when complexes are formed with metal ions. Therefore, the objective of the present study was to evaluate the impact of dietary supplementation of copper acetate (CA), curcumin (CR), and their combination (CA-CR) against Salmonella Typhimurium colonization, intestinal permeability, and cecal microbiota composition in broiler chickens through a laboratory Salmonella infection model. S. Typhimurium recovery was determined on day 10 post-challenge by isolating Salmonella in homogenates of the right cecal tonsil (12 chickens per group) on Xylose Lysine Tergitol-4 (XLT-4) with novobiocin and nalidixic acid. Intestinal integrity was indirectly determined by the fluorometric measurement of fluorescein isothiocyanate dextran (FITC-d) in serum samples from blood obtained on d 10 post-S. Typhimurium challenge. Finally, microbiota analysis was performed using the content of the left caecal tonsil of 5 chickens per group by sequencing V4 region of 16S rRNA gene. RESULTS: The results showed that in two independent studies, all experimental treatments were able to significantly reduce the S. Typhimurium colonization in cecal tonsils (CT, P < 0.0001) compared to the positive control (PC) group. However, only CA-CR was the most effective treatment in reducing S. Typhimurium counts in both independent studies. Furthermore, the serum fluorescein isothiocyanate dextran (FITC-d) concentration in chickens treated with CR was significantly lower when compared to PC (P = 0.0084), which is related to a decrease in intestinal permeability and therefore intestinal integrity. The effect of dietary treatments in reducing Salmonella was further supported by the analysis of 16S rRNA gene sequences using Linear discriminant analysis effect size (LEfSe) since Salmonella was significantly enriched in PC group (LDA score > 2.0 and P < 0.05) compared to other groups. In addition, Coprobacillus, Eubacterium, and Clostridium were significantly higher in the PC group compared to other treatment groups. On the contrary, Fecalibacterium and Enterococcus in CR, unknown genus of Erysipelotrichaceae at CA-CR, and unknown genus of Lachnospiraceae at CA were significantly more abundant respectively. CONCLUSIONS: CR treatment was the most effective treatment to reduce S. Typhimurium intestinal colonization and maintain better intestinal homeostasis which might be achieved through modulation of cecal microbiota.

Evaluation of Ascorbic Acid or Curcumin Formulated in a Solid Dispersion on Salmonella Enteritidis Infection and Intestinal Integrity in Broiler Chickens.

Two experimental models were conducted to evaluate and compare the effect of ascorbic acid (AA) or curcumin formulated in a solid dispersion (SD-CUR) as prophylactic or therapeutic alternatives to prevent or control S. Enteritidis (SE) infection in broiler chickens. In the prophylactic model, dietary administration of AA showed a significant reduction in SE counts in crop compared to the positive control (PC) group (p < 0.05), whereas in cecal tonsils (CT), SD-CUR significantly reduced SE recovery. Superoxide dismutase (SOD) activity was significantly higher in chickens supplemented with AA or SD-CUR, and total intestinal IgA levels were significantly lower in both treatments when compared to the PC group. Serum fluorescein isothiocyanate-dextran (FITC-d) levels were reduced by SD-CUR compared to PC, while AA presented significantly lower total aerobic bacteria. In the therapeutic model, only the dietary administration of AA significantly decreased SE in crop and CT on days 3 and 10 post-challenge. FITC-d levels were significantly lower in both treated groups in comparison to PC, but IgA levels were significantly reduced only by AA. The results suggest that dietary AA and SD-CUR have different modes of action to reduce SE intestinal colonization in two different challenge models in broiler chickens.

Evaluation of the Dietary Supplementation of a Formulation Containing Ascorbic Acid and a Solid Dispersion of Curcumin with Boric Acid against Salmonella Enteritidis and Necrotic Enteritis in Broiler Chickens.

Two experiments were conducted to evaluate the effect of the prophylactic or therapeutic administration of a 0.1% mixture containing ascorbic acid and a solid dispersion of curcumin with polyvinylpyrrolidone and boric acid (AA-CUR/PVP-BA) against Salmonella Enteritidis (S. Enteritidis) in broiler chickens. A third experiment was conducted to evaluate the impact of the dietary administration of 0.1% AA-CUR/PVP-BA in a necrotic enteritis (NE) model in broiler chickens. The prophylactic administration of 0.1% AA-CUR/PVP-BA significantly decreased S. Enteritidis colonization in cecal tonsils (CT) when compared to the positive control group (PC, p < 0.05). The therapeutic administration of 0.1% AA-CUR/PVP-BA significantly reduced the concentration of S. Enteritidis by 2.05 and 2.71 log in crop and CT, respectively, when compared with the PC on day 10 post-S. Enteritidis challenge. Furthermore, the serum FITC-d concentration and total intestinal IgA levels were also significantly lower in chickens that received 0.1% AA-CUR/PVP-BA. Contrary, the PC group showed significantly higher total intestinal IgA levels compared to the negative control or AA-CUR/PVP-BA groups in the NE model. However, 0.1% AA-CUR/PVP-BA showed a better effect in reducing the concentration of S. Enteritidis when compared to the NE model. Further studies with higher concentration of AA-CUR/PVP-BA into the feed to extend these preliminary results are currently being evaluated.

Evaluation of the antimicrobial and intestinal integrity properties of boric acid in broiler chickens infected with Salmonella enteritidis: Proof of concept.

The objectives of the present study were to evaluate the antimicrobial effect of boric acid (BA) on Salmonella enteritidis colonization, intestinal permeability, total intestinal IgA levels, and cecal microbiota composition in broiler chickens. For this purpose, sixty day-old-chicks were randomly allocated to one of two groups: 1) non-treated control group and 2) 0.1% (w/w) BA in feed. All chicks were challenged with 104 cfu of S. Enteritidis per bird at 1-d old. On day 3 and 10 post-challenge, samples of crop and ceca-cecal tonsils (CCT) were aseptically collected to evaluate overall aerobic and lactic acid bacteria, as well as S. Enteritidis recovery. Blood and intestinal samples were also collected to evaluate intestinal permeability and total intestinal IgA production, respectively. On day 10 post-challenge, the concentration of S. Enteritidis in crop and CCT significantly decreased (P < .05) by 2.60 log10 and 5.26 log10, respectively, in the BA group compared to the control group. Furthermore, chickens treated with 0.1% BA had a significant reduction in serum fluorescein isothiocyanate dextran (FITC-d) concentration and lower total intestinal IgA levels when compared to the control group. Interestingly, in the microbiota analysis, only the abundance of Actinobacteria phylum was significantly lower in the BA group in comparison with the control group and significant differences in beta diversity were also observed between comparing groups. The results of the present study suggest that BA could maintain intestinal homeostasis, as well as the balance in the microbiota due to its effectiveness in controlling S. Enteritidis infection.

Assessing the Aflatoxin B1 Adsorption Capacity between Biosorbents Using an In Vitro Multicompartmental Model Simulating the Dynamic Conditions in the Gastrointestinal Tract of Poultry.

Experiments were carried out to evaluate the effectiveness of three different biosorbents (banana peel, Pyracantha leaves, and Aloe powder) in removing aflatoxin B1 (AFB1). A noncommercial mycotoxin binder (zeolite) was used as a reference material. A laboratory model that simulated the in vivo conditions of the poultry gastrointestinal tract was utilized to prove the removal efficiency of the biosorbents when added to AFB1-contaminated diet (100 µg/kg). The concentration of AFB1 was determined using antibody-based immunoaffinity column and spectrofluorometry methodologies. Z potential (ζ), point of zero charge (pHpzc), energy-dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD), Fourier transform infrared spectroscopy with attenuated total reflection (FTIR-ATR), and UV-Vis diffuse reflectance spectroscopy (DRS) techniques were used to further characterize the biosorbents. The addition of the biosorbents (1.5%, w/w) to the diet significantly reduced the bioavailability of AFB1 in the intestinal section. The highest aflatoxin adsorption values were 69% and 70% using Aloe powder and zeolite, respectively. A moderate biosorption uptake of 46% was achieved using Pyracantha leaves. The biomaterial with the lowest removal capacity was banana peel (28%). In conclusion, Aloe powder could be used as an alternative to conventional systems for AFB1 removal.