scholarly journals RESEARCH NOTE: Modified serum fluorescein isothiocyanate dextran (FITC-d) assay procedure to determine intestinal permeability in poultry fed diets high in natural or synthetic pigments

2021 ◽  
pp. 101138
Author(s):  
Christine N. Vuong ◽  
Garrett J. Mullenix ◽  
Michael T. Kidd ◽  
Walter G. Bottje ◽  
Billy M. Hargis ◽  
...  
Keyword(s):  
Intestinal Permeability ◽  
Fluorescein Isothiocyanate ◽  
Research Note ◽  
Assay Procedure ◽  
Fluorescein Isothiocyanate Dextran ◽  
Synthetic Pigments

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 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.

The Effects of External Salinity on the Drinking Rates of the Larvae of Herring, Plaice and Cod

1988 ◽  
Vol 138 (1) ◽  
pp. 1-15 ◽  
Author(s):  
P. TYTLER ◽  
J. H. BLAXTER
Keyword(s):  
Time Course ◽  
Gadus Morhua ◽  
Sea Water ◽  
Fluorescein Isothiocyanate ◽  
Yolk Sac ◽  
Clupea Harengus ◽  
Larval Stages ◽  
First Feeding ◽  
Fluorescein Isothiocyanate Dextran ◽  
External Salinity

Drinking responses to salinity change in the larvae of herring (Clupea harengus L.), plaice (Pleuronectes platessa L.) and cod (Gadus morhua L.) were measured from the time course of uptake of dextran labelled with tritium, following immersion in solutions of 32‰ and 16‰ sea water. The yolk sac and first feeding larval stages of all three species drink in both salinities. Furthermore, post-yolk sac stages appear to adjust their drinking rates to compensate for different salinities in a manner similar to that of the adults. Drinking rates in 32‰ sea water are approximately double those in 16‰. Mass-related drinking rates of larvae are higher than those in adults, but the differences do not match the differences in surface area to mass ratios, suggesting that larval skin is less permeable to water than is adult gill epithelium. Water absorption is indicated by the evidence of concentration of dextran in the gut. The estimates of drinking rates from tritiated dextran uptake are supported by epifluorescence microscopical measurements of the uptake of fluorescein isothiocyanate dextran.

Effect of monoacylglycerol lipase inhibition on intestinal permeability in chronic stress model

2020 ◽  
Author(s):  
Jing Wang ◽  
Xiaohua Zhang ◽  
Chongmei Yang ◽  
Shulei Zhao
Keyword(s):  
Tight Junction ◽  
Chronic Stress ◽  
Intestinal Permeability ◽  
Cannabinoid Receptor ◽  
Fluorescein Isothiocyanate ◽  
Control Group ◽  
Monoacylglycerol Lipase ◽  
Associated Proteins ◽  
Cb1 Antagonist ◽  
Novel Target

Abstract Background: The endocannabinoid 2-arachidonoylglycerol (2-AG) is an anti-nociceptive lipid, which is inactivated through cellular uptake and subsequent catabolism by monoacylglycerol lipase (MAGL). The present study aimed to explore the effects of inhibition of MAGL on intestinal permeability. Methods: We first tested it in differentiated CaCO2 cells after 21 days’ culture. The rat model of water avoidance stress (WAS) was established, and rats were divided into four groups according to intervention. Rats received intraperitoneal injection (i.p.) of an MAGL inhibitor (JZL184) alone, JZL184 and the cannabinoid receptor 1 (CB1) antagonist (SR141716A), JZL184 and a cannabinoid receptor 2 (CB2) antagonist (AM630) or vehicle alone (control). We analyzed the fluorescein isothiocyanate-dextran (FD4) permeability and 2-AG level. Expression of MAGL and tight-junction-associated proteins were detected by western blot. Results: Compared with the control group, MAGL expression was higher and 2-AG levels lower among WAS rats. Intestinal permeability was increased following administration of JZL184 which occurred due to up-regulation of tight-junction-associated proteins Claudin-1, Claudin-2, Claudin-5 and Occludin.Conclusion: The effects of MAGL inhibition were mediated by CB1, indicating that MAGL may represent a novel target for the treatment of reduced intestinal permeability in the context of chronic stress.

NK1 receptors mediate tachykinin-induced increase in microvascular clearance in hamster cheek pouch

1993 ◽  
Vol 265 (2) ◽  
pp. H593-H598
Author(s):  
X. P. Gao ◽  
R. A. Robbins ◽  
R. M. Snider ◽  
J. Lowe ◽  
S. I. Rennard ◽  
...  
Keyword(s):  
Substance P ◽  
Fluorescein Isothiocyanate ◽  
Cheek Pouch ◽  
Neurokinin A ◽  
Plasma Extravasation ◽  
Hamster Cheek Pouch ◽  
Neurokinin B ◽  
Dextran 70 ◽  
Fluorescein Isothiocyanate Dextran ◽  
Neurogenic Plasma Extravasation

The purpose of this study was to determine the receptor subtype(s) that mediates tachykinin-induced neurogenic plasma extravasation in the hamster cheek pouch. Changes in microvascular clearance were quantified by counting the number of leaky sites and calculating the clearance of fluorescein isothiocyanate-dextran [mol wt 70,000 (Dextran 70)] during suffusion of the cheek pouch with substance P, neurokinin A, neurokinin B, and capsaicin. Suffusion of substance P, capsaicin, and neurokinin A, but not neurokinin B, was associated with a significant concentration-dependent increase in leaky site formation and clearance of fluorescein isothiocyanate-Dextran 70 (P < 0.05). However, the responses to substance P and capsaicin were significantly greater than those to neurokinin A. Pretreatment with the selective, nonpeptide NK1 receptor antagonist, CP-96,345, significantly attenuated substance P- and capsaicin-induced but not neurokinin A-induced responses (P < 0.05). These effects were specific, since the 2R,3R enantiomer, CP-96,344, was inactive, and CP-96,345 had no significant effect on adenosine-induced responses. We conclude that, in the hamster cheek pouch, NK1 receptors are the predominant receptors that mediate neurogenic plasma extravasation.

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