In the broiler industry, chicks are often deprived of feed and water up to 48 h posthatch. This delayed access to feed (DAF) has been found to inhibit small intestinal development, compromising growth of the chick. To further understand the impact of DAF on small intestines at the molecular level, many developmental genes that regulate intestinal development were investigated. The objective of this study was to determine the effect of DAF on early posthatch broiler small intestinal morphology, which includes villus height (VH) and crypt depth (CD), and to quantify changes in regulatory genes, such as Olfactomedin 4 (Olfm4), Marker of Ki-67 (Ki-67), Peptide Transporter 1 (PepT1), and Mucin 2 (Muc2), in response to DAF. The Olfm4 mRNA can clearly identify stem cells in the intestinal crypt, which allows VH and CD to be measured, while Ki-67 marks the proliferating cells. The peptide transporter PepT1 is located on intestinal epithelial cells and plays a critical role in transporting di- and tripeptides. Muc2, which is secreted from goblet cells, forms mucus that lines the intestinal epithelial cells acting as a layer of protective coating. Cobb 500 chicks, hatching within a 12 h window, were randomly allocated into three experimental groups: control with no feed delay (ND), 24 h feed delay (D24), and 36 h feed delay (D36). Quantification of Olfm4, Ki-67, PepT1, and Muc2 mRNA abundance were investigated by quantitative PCR, in duodenum, jejunum, and ileum at 0 h, 24 h, 36 h, 72 h, 120 h, and 168 h posthatch. Additionally, localization of cells expressing each gene was visualized using in-situ hybridization at all listed times except 168 h posthatch. Statistical analysis was performed using JMP Pro 14, and significant differences between treatments within a collection day were determined by t-test and one-way ANOVA (P < 0.05). In the ND group, duodenal CD at 0 h was greatest compared to all other time points. With DAF, the duodenal VH of D36 chicks was lower at 36 h (P < 0.001) and 72 h (P = 0.002) compared to ND chicks. In the jejunum and ileum, the VH of D36 chicks was lower at 120 h (P = 0.005) and 72 h (P = 0.03), respectively, compared to ND chicks. In contrast, the VH of D24 chicks at 24 h was greater than ND (P = 0.004) in the jejunum. There was no difference between treatments by 168 h in all intestinal segments. The CD was also lower in DAF groups compared to ND but only in the jejunum and ileum. In contrast, duodenal CD was greater in D24 chicks at 24 h (P = 0.039) and in D36 chicks at 36 h (P < 0.0001) compared to ND chicks, but the difference was no longer significant by 72 h. The VH/CD ratio was lower in all three segments, except the ileum displayed a greater VH/CD ratio in D24 and D36 chicks at 24 h and 36 h, respectively, compared to ND chicks. The mRNA abundance of Olfm4 and Ki-67 was greater in DAF groups upon refeeding, but not until 120 h. The PepT1 mRNA abundance was greater in DAF groups while the abundance of Muc2 mRNA was lower. This difference in mRNA abundance level was more prominent in the duodenum and jejunum. From the analysis of number and distribution of goblet cells found in the upper half and lower half of the villi, expressed as a ratio (VU/VL), a greater ratio was observed in delayed groups compared to ND. In summary, while DAF resulted in altered small intestinal morphology with an effect more pronounced in D36 than D24 chicks, upon refeeding, some genes important to intestinal development were upregulated as a response to the treatment. / Master of Science / In the broiler industry, chicks are often deprived of feed and water up to 48 h posthatch. This delayed access to feed (DAF) was found to negatively impact small intestinal development, compromising their growth. The objective of this study was to determine the effect of DAF on early posthatch broiler small intestinal morphology and to observe the changes in regulatory genes, such as the stem cell marker, proliferating cell marker, absorptive cell marker, and mucus producing cell marker, in response to the DAF. To simulate the DAF condition in the broiler industry, chicks with no feed delay (ND) were tested against chicks with DAF for 24 h (D24) and 36 h (D36). Quantification and cell localization of these cell markers were investigated in the small intestines at early posthatch. In general, DAF chicks had lowered intestinal villus height and crypt depth compared to ND chicks. The mRNA abundance of markers for stem cells and proliferating cells were greater in DAF groups upon refeeding. The mRNA abundance of markers for absorptive cells was greater in DAF groups while the mRNA abundance of markers for mucus producing cells was lowered as a result of DAF. In summary, DAF negatively impacted small intestinal morphology and altered the regulation of some developmental genes important to early posthatch chick performance.

Prior work using a contractility bioassay determined that the serotonin (5-HT) receptor subtype 5-HT2A is present in bovine lateral saphenous veins and plays a role in ergot alkaloid-induced vascular contraction in steers grazing endophyte-infected (Epichloë coenophiala) tall fescue (Lolium arundinaceum). A study was conducted to determine what 5-HT receptors are involved in vasoconstriction of bovine gut vasculature. The findings of this study indicate that 5-HT2A is present and may play a role in ergot alkaloid induced vasoconstriction. A second study was conducted to determine if ergot alkaloids were transported in the small intestine. The active transporter, peptide transporter 1 (PepT1), was evaluated for its role in the transport of various concentrations of ergot alkaloids across Caco-2 cell monolayers. Results indicate that CEPH, ERT, EXT, and LSA do move across Caco-2 cell monolayers, but appear to utilize PepT1 at larger concentrations. Overall, the demonstrated presence of 5-HT2A receptors in the bovine gut vasculature established a potential for vascular interference by ergot alkaloids entering the bloodstream through transepithelial absorption.

Many elasmobranchs are considered top predators with worldwide distribution, and in general these fish play an important role in the transfer of energy from the lower to the upper trophic levels within the marine ecosystem. Despite this, little research has been done regarding the rates of prey ingestion, digestion, and the processes of energy and nutrient absorption. Specifically understudied is enzymatic digestion within the intestinal brush border, which functions to break down macromolecules into smaller subunits for luminal absorption across the gastrointestinal epithelium. Given their carnivorous diet, the present study sought to expand knowledge on nutrient intake in elasmobranchs by focusing on the uptake of products of protein metabolism. To accomplish this, sequence encoding Peptide Transporter 1 (PepT1), a protein found within the brush border membrane (BBM) of higher vertebrates that is responsible for the translocation and absorption of small peptides released during digestion by luminal and membrane-bound proteases, was molecularly identified in the bonnethead shark (Sphyrna tiburo) using degenerate primers based on conserved portions of known PEPT1 sequences from other vertebrates. Sequence encoding Peptide Transporter 2 (PepT2) was also isolated from the S. tiburo scroll valve intestine using the same methodology. PepT1 was then localized using immunocytochemistry with rabbit polyclonal anti-rat PEPT1 in the esophagus, stomach, duodenum, scroll valve intestine, rectum, and pancreas. Vesicle studies were used to identify the apparent affinity of the transporter, and to quantify the rate of uptake by its H+-dependent cotransporter properties, using 3H-glycylsarcosine as a model dipeptide. The results of this study provide insight into the rate and properties of food passage within S. tiburo, and can lead to future work on topics such as physiological regulation of protein metabolism and absorption and how it may vary in elasmobranchs that exhibit different feeding strategies.