Browsing by Author "Jia, Meiting"
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- Delayed access to feed affects broiler small intestinal morphology and goblet cell ontogenyLiu, Kuan-Ling; Jia, Meiting; Wong, Eric A. (2020-11)Broilers are often deprived of feed and water for up to 48 h after hatch. This delayed access to feed (DAF) can inhibit small intestine development. The objective of this study was to determine the effects of DAF on small intestinal morphology, mRNA abundance of the goblet cell marker Muc2 and absorptive cell marker PepT1, and the distribution of goblet cells in young broilers. Cobb 500 chicks, hatching within a 12-h window, were randomly allocated into 3 groups: control with no feed delay (ND), 24-h feed delay (DAF24), and 36-h feed delay (DAF36). Morphology, gene expression, and in situ hybridization analyses were conducted on the duodenum, jejunum, and ileum at 0, 24, 36, 72, 120, and 168 h after hatch. Statistical analysis was performed using a t test for ND and DAF24 at 24 h. A 2-way ANOVA and Tukey's HSD test (P < 0.05) were used for ND, DAF24, and DAF36 from 36 h. At 24 to 36 h, DAF decreased the ratio of villus height/crypt depth (VH/CD) in the duodenum but increased VH/CD in the ileum due to changes in CD, whereas at 72 h, DAF decreased VH/CD due to a decrease in VH. The mRNA abundance of PepT1 was upregulated, while Muc2 mRNA was downregulated in DAF chicks. Cells expressing Muc2 mRNA were present along the villi and in the crypts. The ratio of the number of goblet cells found in the upper half to the lower half of the villus was greater in DAF chicks than in ND chicks, suggesting that DAF affected the appearance of new goblet cells. The number of Muc2 mRNA-expressing cells in the crypt, however, was generally not affected by DAF. In conclusion, DAF transiently affected small intestinal morphology, upregulated PepT1 mRNA, downregulated Muc2 mRNA, and changed the distribution of goblet cells in the villi. By 168 h, however, these parameters were not different between ND, DAF24, and DAF36 chicks.
- Effect of probiotics or high incubation temperature on gene expression and cell organization of the small intestine and yolk sac of chicksJia, Meiting (Virginia Tech, 2021-11-30)The small intestine and yolk sac (YS) are important organs for nutrient absorption and innate immunity in chickens during the post-hatch or prehatch periods. These organs share a similar structure of epithelial cell-lined villi with tight junctions between adjacent cells. Probiotics have been reported to improve chicken growth performance and gut health including promotion of intestinal morphology. However, there are few studies that show the effect of probiotics on ontogeny of intestinal epithelial cells and antimicrobial peptides, or intestinal integrity in young healthy chicks. Heat stress during incubation was shown to increase mortality and decrease hatchability of chicks, while no studies have investigated the effect of heat stress on the integrity of the YS, which might be related to hatching performance. There were four studies conducted in this research: 1) a comparison of the effect of two probiotics on the ontogeny of small intestinal epithelial cells in young chicks; 2) the effect of two probiotics on mRNA abundance of tight junction proteins in the small intestine of young chicks; 3) the effect of high incubation temperature on mRNA abundance of tight junction proteins in the YS of broiler embryos; and 4) comparison of avian defense peptide mRNA abundance in the YS of broilers and layers. In study 1, Probiotics transiently decreased body weight gain (BWG) from day 2 to day 4, but did not affect body weight (BW) from day 2 to day 8, and small intestinal weight and intestinal morphology from day 2 to day 6. Probiotics did not affect marker gene expression of intestinal stem cells (Olfm4) and goblet cells (Muc2) in all small intestinal segments, but did increase expression of a marker gene of proliferating cells (Ki67), and decreased an antimicrobial peptide (liver-enriched antimicrobial peptide 2, LEAP2) in the jejunum at day 4. Probiotic 1 decreased PepT1, a marker of enterocytes in the duodenum at day 4. These results suggest that probiotics did not improve growth performance and intestinal morphology in young healthy chicks, but temporarily promoted intestinal epithelial cell proliferation and decreased LEAP2 antimicrobial peptide expression in the jejunum. In situ hybridization (ISH) showed that Ki67+ proliferating cells were mainly located in the crypt region and the blood vessels of villi. In study 2, Probiotic supplementation to newly hatched chicks for less than one week did not affect mRNA abundance of the tight junction proteins in the small intestine. Occludin (OCLN) mRNA, which was detected by ISH to be expressed in intestinal epithelial cells in both the villus and crypt regions, was greater in the duodenum of female chicks than males. In study 3, high incubation temperature starting from embryonic day 12 (E12) affected mRNA abundance of the tight junction proteins in the YS, including increased zonula occluden 1 (ZO1) at E13, increased junctional adhesion molecule A (JAMA) and heat shock protein 90 (HSP90) at E17, but decreased tight junction protein JAMA at E19 and OCLN at day of hatch (DOH). These results showed that the YS tight junction proteins were increased by short term heat exposure but decreased by long term heat exposure. In study 4, the expression of avian β defensin 10 (AvBD10), CATHs and toll-like receptors in the YS was examined. Toll-like receptors were highly expressed in the YS at early incubation stages (E7), while CATHs showed a peak expression from E9 to E13, which was similar to the expression pattern of AvBD10. CATHs and AvBD10 mRNA temporal expression patterns were similar in broilers and layers, while their expression levels were different. Layers, especially brown layers, had greater mRNA abundance for antimicrobial peptides such as AvBD10, CATH1, and CATH2 in the YS. These results demonstrate that the antimicrobial peptide temporal expression patterns in the YS are not affected by breed, but their expression levels are affected by breed. In summary, the small intestine and the YS are essential for nutrient uptake, innate immunity, and maintenance of integrity. The ontogeny of intestinal epithelial cells, such as proliferating cells can be modulated by probiotic supplementation. Similar to the small intestine, the YS can also express tight junction proteins, which can be affected by high incubation temperature. Antimicrobial peptide expression in the intestine of healthy young chicks is also transiently decreased by probiotic supplements. Avian defensin and cathelicidin expression patterns in the YS were not affected by breed.
- Research Note: Intestinal morphology and gene expression changes in broilers supplemented with lysolecithinCloft, Sara E.; Jia, Meiting; Wong, Eric A. (Elsevier, 2021-07-01)Lysolecithin is used as a feed additive to aid fat digestion and absorption in broiler chickens. Previous research has shown that dietary fat source influences how broilers respond to lysolecithin supplementation. Therefore, the objective of this study was to investigate the effect of lysolecithin on a diet formulated with soybean oil on jejunum morphology and expression of selected genes in broiler chickens. Male Cobb 500 chickens were fed a Control diet or the Control diet supplemented with lysolecithin (TRT) from day of hatch to day 28. Jejunal samples were collected at day 10 for morphological and gene expression analysis. Feeding the TRT diet did not affect BW, villus height (VH), crypt depth (CD) or VH/CD ratio compared to Control fed chickens. Differential gene expression in the jejunum was analyzed using a custom microarray. Using a t test, 36 genes were found to be upregulated in TRT fed chickens compared to chickens fed the Control diet. The two most upregulated genes were carbonic anhydrase VII and interleukin 8-like 2, which are associated with healthy intestines. In summary, lysolecithin supplementation in a diet formulated with soybean oil caused no morphological changes but upregulated a number of genes in the jejunum.
- Temporal expression of avian β defensin 10 and cathelicidins in the yolk sac tissue of broiler and layer embryosJia, Meiting; Fulton, J. E.; Wong, Eric A. (Elsevier, 2023-02-01)The yolk sac is a multifunctional organ, which not only participates in nutrient absorption, but also plays an important role in immune function. The objective of this study was to compare the mRNA abundance of avian β-defensin 10 (AvBD10) and 3 cathelicidins (CATH1, CATH2, and CATH3) in the yolk sac tissue (YST) of commercial broilers and white egg and brown egg commercial layers. AvBD10 and CATH mRNA abundance was analyzed using two-way ANOVA and Tukey's test, with P < 0.05 being considered significant. AvBD10 and CATH mRNA showed similar temporal expression patterns in the YST of both broiler and layers, with an increase from embryonic day (E) 7 to E9 through E13 followed by a decrease to day of hatch. AvBD10 mRNA showed a breed × age interaction with greater expression in the YST of both layers compared to broilers at E9 and E11. CATH1 mRNA was greater in the YST of brown egg layers than broilers. CATH2 mRNA showed a breed × age interaction, with greater expression in the YST of brown egg layers than broilers at E11. CATH3 mRNA showed no difference in the YST between layers and broilers. Because broilers and brown egg layers are genetically related, these results show that selection for production parameters (broiler vs. layer) and not genetic relatedness (white egg layer vs. brown egg layer and broilers) is the basis for the differences in AvBD10, CATH1, and CATH2 mRNA in the YST of broilers and layers. The yolk-free body weights of broiler embryos were greater than that of both brown and white egg layers from E9 to 17. One possible explanation is that the reduced expression of AvBD10, CATH1 and CATH2 mRNA in the YST of broilers compared to layers at E9 and 11 may be due to faster embryonic growth at the expense of host defense peptide expression in broilers compared to layers.