scholarly journals A Microencapsulated Mixture of Eugenol and Garlic Tincture Supplementation Mitigates the Effect of Necrotic Enteritis on Intestinal Integrity and Increases Goblet Cells in Broilers

2021 ◽  
Vol 9 (7) ◽  
pp. 1451
Author(s):  
Alip Kumar ◽  
Sarbast K. Kheravii ◽  
Catherine Ionescu ◽  
Alexandra Blanchard ◽  
Reza Barekatain ◽  
...  

This study was conducted to examine the effects of a plant extract mixture, a microencapsulated product composed of eugenol and garlic tincture (PE), on intestinal health in broilers under necrotic enteritis (NE) challenge. A total of 960 d-old mixed-sex Cobb 500 chicks were randomly distributed to 48-floor pens housing 20 birds per pen. Six treatments were applied: UC, unchallenged control; CC, challenged control; PE, challenged group plus PE; AM, challenged group plus antimicrobial (AM); FAP, challenged group plus a full dose of AM with PE; HAP, challenged group plus a half dose of AM with PE in starter, grower and finisher phases. Birds in the challenged groups were inoculated with Eimeria spp. on d 9 and Clostridium perfringens on d14. On d 16, the CC group had increased serum fluorescein isothiocyanate dextran (FITC-d), reduced villus surface area, goblet cell number, upregulated CLDN1, JAM2 genes and reduced microbial diversity compared to the UC group (p < 0.05). Birds fed PE had reduced FITC-d, increased goblet cell number and Bifidobacterium compared to the CC group (p < 0.05). Birds fed PE had reduced CLDN5 expression in male birds, and Bacteroides spp. in female birds than CC group (p < 0.05). These findings suggest that PE supplementation mitigates the effect of NE by improving the intestinal health of birds.

Animals ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1432
Author(s):  
Alip Kumar ◽  
Sarbast K. Kheravii ◽  
Lily Li ◽  
Shu-Biao Wu

This study evaluated the potential of monoglyceride blend (MG) and buffered formic acid (FA) as alternatives to antibiotics in the performance and intestinal health of broilers under clinical necrotic enteritis (NE) challenge. A total of 544 as-hatched Ross 308 broiler chicks were randomly distributed to 32-floor pens housing 17 birds per pen. The four treatments were: NC—non-additive control; ZBS—antibiotic group supplemented with zinc bacitracin and salinomycin; MG—additive MG supplementation in the starter phase only; and MGFA—additive MG in starter phase and FA in grower and finisher phases. All birds were challenged with Eimeria spp. and Clostridium perfringens. Results showed that the NC group had lower BWG and higher FCR than the ZBS group in the grower and overall period (p < 0.05). The NC group had higher NE-caused mortality (days 14 to 17) than the ZBS group (p < 0.05). Birds fed MG had lower NE-caused mortality than the NC group (p < 0.05). Birds fed MG had upregulated jejunal tight junction protein1 (TJP1) and immunoglobulin (IgG) on day 16 and improved gross energy digestibility on day 24 than the NC group (p < 0.05). These findings suggest that supplementation of MG may improve intestinal health and protect birds from clinical NE occurrence.


2020 ◽  
Author(s):  
Anaisa A. Leyva-Diaz ◽  
Daniel Hernandez-Patlan ◽  
Bruno Solis-Cruz ◽  
Bishnu Adhikari ◽  
Young Min Kwon ◽  
...  

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.


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Anaisa A. Leyva-Diaz ◽  
Daniel Hernandez-Patlan ◽  
Bruno Solis-Cruz ◽  
Bishnu Adhikari ◽  
Young Min Kwon ◽  
...  

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.


2020 ◽  
Author(s):  
Anaisa A. Leyva-Diaz ◽  
Daniel Hernandez-Patlan ◽  
Bruno Solis-Cruz ◽  
Bishnu Adhikari ◽  
Young Min Kwon ◽  
...  

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.


2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 114-114
Author(s):  
Bonjin Koo ◽  
Chengbo Yang ◽  
Martin Nyachoti

Abstract The aim was to investigate the effects of dietary Met and Cys supplementation in immunologically challenged-weaned pigs. Forty weaned piglets (6.51 ± 0.29 kg) were randomly allocated to five treatments and fed the assigned diets for 9 days. The treatments included: (1) sham-challenged control (SCC), (2) challenged control (CC), (3) MET (CC + 0.1% DL-Met supplementation), (4) CYS (CC + 0.1% L-Cys supplementation), and (5) MET+CYS (CC + 0.1% DL-Met and 0.1% L-Cys supplementation). Piglets were intramuscularly injected with either lipopolysaccharides (LPS; 30 μg/kg) or saline solution on day 7. All data were analyzed using single degree of freedom contrasts in the mixed procedure of SAS to test the effect of LPS challenge, Met supplementation, Cys supplementation, and Met × Cys supplementation. The CC group had lower (P &lt; 0.05) villi height and villi surface area and glutathione (GSH) concentration and higher (P &lt; 0.05) flux of 4-kDa fluorescein isothiocyanate dextran (FD4) in the jejunum than SCC group. Dietary Cys supplementation increased (P &lt; 0.05) villi surface area, GSH concentration and reduced GSH: oxidized GSH and decreased (P &lt; 0.05) the flux of FD4 in the jejunum. Dietary Met supplementation enhanced (P &lt; 0.05) hepatic GSH content. Pigs in the MET group had greater mRNA abundance of IL-6 and IL-10 (P &lt; 0.05) and lower serum IL-8 concentration than those in the CC group. There were Met × Cys interactions (P &lt; 0.05) in serum IL-4 and IL-8 concentrations and hepatic Trolox equivalent antioxidant capacity. In conclusion, dietary L-Cys supplementation restored intestinal integrity and GSH concentration that were affected by LPS challenge. Dietary DL-Met supplementation improved hepatic GSH and reduced systemic inflammatory response, but antagonistic interaction with dietary Cys supplementation was observed in inflammatory response and redox status.


2021 ◽  
Vol 4 (Supplement_1) ◽  
pp. 6-7
Author(s):  
E Fekete ◽  
C B Amat ◽  
T Allain ◽  
M Hollenberg ◽  
K Mihara ◽  
...  

Abstract Background Giardia duodenalis has been shown to alter the structure of the intestinal mucus layers during infection via obscure mechanisms. We hypothesize that goblet cell activity may be disrupted in part due to proteolytic activation of protease-activated receptor 2 (PAR2) by Giardia proteases, resulting in disruption of mucus production and secretion by intestinal goblet cells. Aims Characterize alterations in goblet cell activity during Giardia infection, focusing on the roles of Giardia protease activity and PAR2. Methods Chinese hamster ovary cells transfected with nano-luciferase tagged PAR2 were incubated with Giardia NF or GSM trophozoites. Cleavage within the activation domain results in release of enzymes into the supernatant. Luminescence in the supernatant was measured as an indication of PAR cleavage by Giardia. LS174T, a human colonic mucus-producing cell line, was infected with Giardia trophozoites (isolates NF, WB, S2, and GSM). Prior to infection, trophozoites were treated with E64, a broad-spectrum cysteine protease inhibitor, and LS174T were treated with a PAR2 antagonist, a calcium chelator, or an ERK1/2 inhibitor. Quantitative PCR (qPCR) was performed for the MUC2 mucin gene. Wild-type (WT) and PAR2 knockout (KO) mice were infected with Giardia. Colonic mucus was stained using fluorescein-coupled wheat-germ agglutinin (WGA), and qPCR was performed for Muc2 and Muc5ac. Results Giardia trophozoites cleaved PAR2 within the N-terminal activation domain in a cysteine protease-dependent manner. Cleavage was isolate dependent, with isolates that show higher protease activity cleaving at a higher rate. High protease activity Giardia isolates increased MUC2 gene expression in LS714T. This increase was attenuated by inhibition of Giardia cysteine protease activity, and by antagonism of PAR2, inhibition of calcium release, or inhibition of ERK1/2 activity in LS174T cells. Both Muc2 and Muc5ac expression were upregulated in the colons of WT mice in response to Giardia infection, while in the jejunum Muc2 expression decreased and Muc5ac expression increased. In KO, no changes in gene expression were seen in the colon in response to Giardia infection, while in the jejunum, Muc2 expression was unchanged and Muc5ac expression decreased. Both WT infected and KO noninfected mice showed thinning of the colonic mucus layer compared to WT controls. There was some recovery in thickness in KO infected mice. Conclusions PAR2 plays a significant role in the regulation of mucin gene expression in mice and in a human colonic cell line. Results suggest that Giardia cysteine proteases cleave and activate PAR2, leading to calcium release and activation of the MAPK pathway in goblet cells, ultimately leading to altered mucin gene expression. Findings identify a novel regulatory pathway for mucus production by intestinal goblet cells. Funding Agencies CAG, CCC


Animals ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 2213
Author(s):  
Jared Ruff ◽  
Thaina L. Barros ◽  
Joy Campbell ◽  
Ricardo González-Esquerra ◽  
Christine N. Vuong ◽  
...  

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.


Membranes ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 550
Author(s):  
Xiaofan Du ◽  
Jing Wang ◽  
Lan Chen ◽  
Zhenxi Zhang ◽  
Cuiping Yao

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%.


1995 ◽  
Vol 311 (1) ◽  
pp. 293-297 ◽  
Author(s):  
M Tomita ◽  
H Itoh ◽  
N Ishikawa ◽  
A Higa ◽  
H Ide ◽  
...  

A cDNA encoding mouse intestinal trefoil factor (mITF) was successfully cloned and sequenced from the small intestine of C57BL/6 mouse by using the combination of reverse transcription-PCR and rapid amplification of cDNA ends methods. The gene was, similar to rat and human ITFs, mainly expressed in the small and large intestine. The mITF expression was up-regulated during the recovery phase after depletion of goblet cells in acetic acid-induced colitis. On the other hand, the expression in the jejunum was not altered, while goblet cell hyperplasia was induced by Nippostrongylus brasiliensis infection. These results suggest that the mITF expression did not simply correlate with the number of goblet cells. The mITF may play an important role in the maintenance and repair of mucosal function of the rectum. Additionally, the mITF in the jejunum may play a role in alteration of the physicochemical nature of goblet cell mucins, thereby affecting the establishment of intestinal helminths.


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