scholarly journals Assay considerations for fluorescein isothiocyanate-dextran (FITC-d): An indicator of intestinal permeability in broiler chickens

2021 ◽  
pp. 101202
Author(s):  
Jundi Liu ◽  
Po-Yun Teng ◽  
Woo K. Kim ◽  
Todd J. Applegate
Keyword(s):  
Intestinal Permeability ◽  
Broiler Chickens ◽  
Fluorescein Isothiocyanate ◽  
Fluorescein Isothiocyanate Dextran

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

2020 ◽  
Author(s):  
Anaisa A. Leyva-Diaz ◽  
Daniel Hernandez-Patlan ◽  
Bruno Solis-Cruz ◽  
Bishnu Adhikari ◽  
Young Min Kwon ◽  
...  
Keyword(s):  
16S Rrna ◽  
Intestinal Permeability ◽  
Broiler Chickens ◽  
Copper Acetate ◽  
Fluorescein Isothiocyanate ◽  
Rrna Gene ◽  
Microbiota Composition ◽  
Intestinal Integrity ◽  
Fluorescein Isothiocyanate Dextran ◽  
Cecal Microbiota

Abstract 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 day 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 the 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.

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

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Anaisa A. Leyva-Diaz ◽  
Daniel Hernandez-Patlan ◽  
Bruno Solis-Cruz ◽  
Bishnu Adhikari ◽  
Young Min Kwon ◽  
...  
Keyword(s):  
16S Rrna ◽  
Intestinal Permeability ◽  
Broiler Chickens ◽  
Copper Acetate ◽  
Fluorescein Isothiocyanate ◽  
Rrna Gene ◽  
Microbiota Composition ◽  
Intestinal Integrity ◽  
Fluorescein Isothiocyanate Dextran ◽  
Cecal Microbiota

Abstract 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.

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

2020 ◽  
Author(s):  
Anaisa A. Leyva-Diaz ◽  
Daniel Hernandez-Patlan ◽  
Bruno Solis-Cruz ◽  
Bishnu Adhikari ◽  
Young Min Kwon ◽  
...  
Keyword(s):  
16S Rrna ◽  
Intestinal Permeability ◽  
Broiler Chickens ◽  
Copper Acetate ◽  
Fluorescein Isothiocyanate ◽  
Rrna Gene ◽  
Microbiota Composition ◽  
Intestinal Integrity ◽  
Fluorescein Isothiocyanate Dextran ◽  
Cecal Microbiota

Abstract 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 day 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 the 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.

scholarly journals Evaluation of Curcumin and Copper Acetate Against Salmonella Typhimurium Infection, Intestinal Permeability, and Cecal Microbiota Composition in Broiler Chickens

2020 ◽  
Author(s):  
Anaisa A. Leyva-Diaz ◽  
Daniel Hernandez-Patlan ◽  
Bruno Solis-Cruz ◽  
Bishnu Adhikari ◽  
Young Min Kwon ◽  
...  
Keyword(s):  
Intestinal Permeability ◽  
Broiler Chickens ◽  
Bacterial Resistance ◽  
Copper Acetate ◽  
Fluorescein Isothiocyanate ◽  
Feed Additives ◽  
Microbiota Composition ◽  
Linear Discriminant ◽  
Cecal Microbiota ◽  
The Impact

Abstract 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.Results: The obtained results showed that in two independent studies, all experimental treatments were able to significantly reduce the S. Typhimurium colonization in cecal tonsils (CT, p < 0.05) compared to the positive control (PC) group. However, CR and CA-CR were the best treatments reducing S. Typhimurium counts. Furthermore, the serum fluorescein isothiocyanate dextran (FITC-d) concentration in chickens treated with CR was significantly lower when compared to PC (p < 0.05). The effect of dietary treatments in reducing S. Typhimurium colonization was further supported by the Linear discriminant analysis effect size (LEfSe) analysis of microbiome data, where Salmonella was significantly enriched in PC group (LDA score > 2.0 and p<0.05). In addition, Coprobacillus, Eubacterium, and Clostridium were significantly higher in the PC group compared to other treatment groups. On the contrary, Fecalibacterium and Enterococcus at CR, unknown genus of Erysipelotrichaceae at CA-CR, and unknown genus of Lachnospiraceae at CA were significantly abundant respectively.Conclusions: CR treatment was the most effective treatment to reduce S. Typhimurium intestinal colonization and intestinal permeability associated with variations in the cecae microbiota structure in broiler chickens.

scholarly journals Spray-Dried Plasma Improves Body Weight, Intestinal Barrier Function, and Tibia Strength during Experimental Constant Heat Stress Conditions

Animals ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 2213
Author(s):  
Jared Ruff ◽  
Thaina L. Barros ◽  
Joy Campbell ◽  
Ricardo González-Esquerra ◽  
Christine N. Vuong ◽  
...  
Keyword(s):  
Heat Stress ◽  
Intestinal Barrier ◽  
Fluorescein Isothiocyanate ◽  
Serum Levels ◽  
Negative Control ◽  
Intestinal Barrier Function ◽  
Negative Consequences ◽  
Fluorescein Isothiocyanate Dextran ◽  
Prolonged Heat ◽  
Spray Dried

The aim of this study was to see how spray-dried plasma (SDP) supplementation affected broiler chicken performance, intestinal permeability, and bone strength during persistent heat stress. One-day-old chicks (n = 480) were randomly assigned into twelve environmental corrals; four thermoneutral (TN-negative control, maintained at 24 °C from d 21–42); four heat stress (HS, exposed to 35 °C from d 21–42); and four heat stress treated with 2% SDP in the feed until d 28 followed by 1% SDP until d 42 (HS-SDP). The performance and serum levels of fluorescein isothiocyanate-dextran (FITC-d) were evaluated at d 21, 28, 35, and 42. The tibias strength was evaluated on d 21 and 42. The increment in chicken temperature (p < 0.05) was observed two h following the increase in environmental temperature in both HS groups and was associated with decreased performance parameters compared with the TN group. At d 42 of age, the chickens exposed to HS had an impaired gut permeability and decreased tibia strength compared to the TN group (p < 0.05). However, partially feeding SDP mitigated these adverse effects significantly. These findings imply that using SDP strategically during stressful times, such as prolonged heat stress, may help mitigate its negative consequences.

scholarly journals Delivery of Foreign Materials into Adherent Cells by Gold Nanoparticle-Mediated Photoporation

Membranes ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 550
Author(s):  
Xiaofan Du ◽  
Jing Wang ◽  
Lan Chen ◽  
Zhenxi Zhang ◽  
Cuiping Yao
Keyword(s):  
Fluorescein Isothiocyanate ◽  
Adherent Cells ◽  
Thermal Dynamics ◽  
Cervical Cell ◽  
Nanosecond Pulse Laser ◽  
Fluorescein Isothiocyanate Dextran ◽  
Extracellular Materials ◽  
Foreign Materials ◽  
Cell Medium ◽  
Delivery Efficiency

Delivering extracellular materials into adherent cells presents several challenges. A homemade photoporation platform, mediated by gold nanoparticles (AuNPs), was constructed to find a suitable method for finding all adherent cells in this process with high delivery efficiency. The thermal dynamics of AuNPs could be monitored. Based on this system, 60 nm AuNPs were selected to be attached to cells for optimal photoporation. After irradiating the cells covered with AuNPs using a nanosecond pulse laser, fluorescein isothiocyanate-dextran in the medium were delivered into optoporated adherent HeLa (human cervical cell lines) cells. The delivery efficiency and cell viability of this process were evaluated using a fluorescence microscope and flow cytometry. The experimental results showed that targeting cells using antibodies, laser irradiation from the top of the cell culture well, and reducing the cell medium are important for improving the delivery efficiency. The optimal loading efficiency for adherent HeLa cells was 53.4%.

Spreading of microinjected horseradish peroxidase to nondescendant cells in embryos of Patella (Mollusca, Gastropoda)

Development ◽  
1987 ◽  
Vol 100 (4) ◽  
pp. 713-722
Author(s):  
W.M. Kuhtreiber ◽  
F. Serras ◽  
J.A.M. van den Biggelaar
Keyword(s):  
Molecular Weight ◽  
Stem Cell ◽  
Horseradish Peroxidase ◽  
High Molecular Weight ◽  
Fluorescein Isothiocyanate ◽  
Contact Phase ◽  
High Molecular Weight Form ◽  
Contact Stage ◽  
Fluorescein Isothiocyanate Dextran ◽  
Molecular Weight Form

We have injected horseradish peroxidase (HRP) and fluorescein-isothiocyanate dextran (FD) into cells and into the blastocoelic cavity of Patella vulgata embryos, before and during the interval between 5th and 6th cleavage, in which the mesodermal stem cell is determined by means of interactions between the central 3D macromere and the contacting animal micromeres. Intracellular injections of HRP at different stages showed that, whereas before this contact phase no spreading of label was observed, a clear intercellular transfer of HRP was found after the contact was established. Control experiments showed that it was HRP in its intact, high molecular weight form that was transferred in the living embryo. Injections of HRP into the blastocoelic cavity gave essentially the same results. In these cases, the HRP was taken up by the cells from contact stage onwards. When FD was injected into the blastocoelic cavity, no uptake was observed, not even after prolonged presence of FD in it. However, when HRP and FD were mixed, both were taken up, starting at contact stage. Differences in labelling pattern of HRP, as compared with FD, and a shift of the FD fluorescence after uptake, suggest that receptor-mediated endocytosis is involved. The possible morphogenetic significance of the transfer mechanism is discussed.

scholarly journals Intestinal Barrier Function and Performance of Broiler Chickens Fed Additional Arginine, Combination of Arginine and Glutamine or an Amino Acid-Based Solution

Animals ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 2416
Author(s):  
Reza Barekatain ◽  
Tristan Chalvon-Demersay ◽  
Clive McLaughlan ◽  
William Lambert
Keyword(s):  
Amino Acid ◽  
Intestinal Permeability ◽  
Broiler Chickens ◽  
Intestinal Inflammation ◽  
Intestinal Barrier ◽  
Basal Diet ◽  
Intestinal Barrier Function ◽  
Glutathione Synthetase ◽  
Feed Conversion ◽  
Experimental Treatments

Two experiments were conducted to investigate the effect of arginine (Arg); the combination of Arg and glutamine (Gln); as well as an amino acid-based solution (MIX) containing Arg, Gln, threonine (Thr), and grape extract, on performance, intestinal permeability, and expression of selected mechanistic genes. Using 240 male Ross 308 off-sex broiler chickens, four experimental treatments were replicated six times with 10 birds per replicate. The experimental treatments included 5 g/kg Arg, 2.5 g/kg Arg and 2.5 g/kg Gln, and 1 g/kg MIX added to a basal diet as control. In the second study, the four dietary treatments were then given to 24 birds with or without a synthetic glucocorticoid, dexamethasone (DEX), as a gut dysfunction model. Feed conversion ratio was improved by all the supplemented treatments from day 7 to 35 of age (p < 0.001). DEX injections increased (p < 0.001) the intestinal permeability in all treatments, which tended to be reversed by Arg or MIX. Additional Arg, Arg-Gln, and MIX suppressed (p < 0.05) the overexpression of IL-1β generated by DEX. Feeding birds with MIX treatment increased (p < 0.05) expression of SGLT-1 and glutathione synthetase. In conclusion, tested amino acid supplements were effective in improving feed efficiency and restraining intestinal inflammation caused by DEX through IL-1β pathway.

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