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Biomedical subjects

D Sklan

Publications and source records attributed to D Sklan.

At least 37 records · Page 2Linked to original sources

Starving affects CDX gene expression during small intestinal development in the chick.

The small intestine of the chicken undergoes intensive changes in the immediate posthatch period, increasing in size and developing crypts, villi and mature enterocytes. During this time, chicks are also transferring from nutrition based on the lipid-rich yolk to exogenous carbohydrate-rich feeds. The cdx homeobox genes participate in axial patterning and in definition of cell identity in embryos, and some cdx genes remain active postpartum in organs such as the intestine. In this study, the transcription patterns of two of these genes, cdxA and cdxB, were examined in the small intestine of the embryo and posthatch chick; in addition, the effects on these genes of starving for 48 h at hatch were examined. Both cdx transcription factors were upregulated toward the time of hatch and were observed in proliferating enterocytes; this enhanced expression continued posthatch. Distribution of cdxA changed with age and was found at higher concentrations in mature enterocytes. Starving from 0 to 48 h posthatch retarded growth and decreased enterocyte proliferation and expression of cdxA and cdxB. After access to feed, expression of cdx genes was enhanced. Chicken homeobox genes cdxA and cdxB are expressed in all enterocytes during embryonic and posthatch development; however, cdxA may have a role in enterocyte maturation posthatch. CdxB was expressed later in development then previously reported.

Animals↗

Nutrient use in chicks during the first week posthatch.

We examined effects of feeding chicks differing levels of macronutrients on performance during the first 7 d posthatch in this study. Four experiments were conducted using male Ross x Ross broiler chicks. The first experiment examined, in a 3 x 3 factorial design, the effect of feeding fat at 3, 7, and 11% and protein at 18, 23, and 28%. With increasing protein and fat levels feed intake and BW decreased. The second experiment examined, in a 3 x 2 factorial design, the effect of fat at 3, 7, and 10% and cellulose at 3 and 13%. Increasing fat in the diet decreased feed intake at both cellulose levels with little effect on BW. High cellulose decreased feed intake and BW. Experiment 3 examined, in a 3 x 2 factorial design, the effect of protein at 19, 23, and 27% and cellulose at 3 and 13%. Increasing protein levels decreased feed intake with little effect on BW. Cellulose decreased feed intake as levels increased. The fourth experiment examined, in a 4 x 2 factorial, the effect of feeding protein at 19, 21, 23, and 25% and fat at 4.5 and 9%, for 1 wk followed by standard diets through marketing. Significant differences in feed intake, BW and feed efficiency were observed at 7 d, but by 18 d differences were not significant and by 31 and 41 d no treatment effects were observed. Fat and protein percentage in the carcass in all experiments were not altered by dietary treatments and thus the efficiency of protein and fat retention decreased with increasing dietary intake. This study indicates that feeding diets with varying macronutrient levels to chicks during the first week post-hatch has distinct effects as compared to older broilers. It appears that once limiting amino acids and energy are provided the influence of dietary composition on immediate posthatch growth is limited.

Aging↗

Response, tolerance and ignorance following oral exposure to a single dietary protein antigen in Gallus domesticus.

The objective of this investigation was to analyze conditions leading to antibody responses against innocuous dietary protein antigens in the chick. The physical form of antigen was found to be important for immunization of mature chicks: bovine serum albumin (BSA) in solution was a powerful immunogen, while BSA powder was ignored. When BSA was fed to newly hatched chicks, either in solution or as powder, specific oral tolerance was induced. Tolerance increased with the dose of antigen fed, and was most effective in suppressing BSA-specific oral immunization. Hence, immune responses of mature chicks to innocuous dietary proteins are not likely to constitute a health hazard due to (a) prevalence of oral tolerance induced at hatch, and (b) availability of dietary proteins in solid form.

Administration, Oral↗

The effect of fasting at different ages on growth and tissue dynamics in the small intestine of the young chick.

The small intestines of hatching chicks undergo rapid developmental changes in the immediate post-hatch period when the birds are making the transition from endogenous nutrient supply from yolk to dependence on exogenous feed. This transition usually only begins 48 h or more after hatching, owing to logistical considerations of production. The effects of fasting for 48 h at different times during this critical period on small intestinal development and enterocyte dynamics were examined by morphometric determinations and use of staining for proliferative-cell nuclear antigen and 5-bromo-2-deoxyuridine. The effects of fasting were specific to both time of fasting and the intestinal segment examined. Decreased development was found in the duodenum and jejunum, but was less apparent in the ileum. Fasting between 0 and 48 h decreased crypt size in the duodenum and jejunum, the number of crypts per villus, crypt proliferation, villus area and the rate of enterocyte migration. Fasting at later times resulted in smaller effects, although the jejunum appeared to be the most sensitive of the intestinal segments. Growth was correlated with the number of cells in the crypts, the number of cells along the villus and the segment surface area. The common practice whereby feed is first available to chicks more than 48 h post-hatch may depress subsequent development.

Animals↗

Yolk and exogenous feed utilization in the posthatch chick.

This study examined in posthatch chicks the partition of yolk utilization between transport to the circulation and secretion to the intestine and determined absorption and plasma concentrations of some metabolites. The presence of feed in the gastrointestinal tract enhanced yolk secretion to the small intestines. Absorption of glucose, methionine, and oleic acid was determined in vivo close to hatch using 141Ce as a nonabsorbed reference substance. Glucose and methionine uptake was low posthatch and increased with age and feed intake. Oleic acid uptake was high at hatch and changed little with age. Increased intake of lipids depressed the percentage oleic acid absorption. Plasma concentrations of glucose, Na, phospholipids, and nonesterified fatty acids did not change with age. Triiodothyronine and triglyceride concentrations increased with age in the plasma of fed chicks but decreased during feed deprivation. Feed intake stimulated posthatch secretion of yolk to the small intestines and triggered uptake mechanisms for hydrophilic compounds.

Aging↗

Changes in growth and function of chick small intestine epithelium due to early thermal conditioning.

The effect of exposure to heat at 3 d of age on small intestine functionality and development was assayed by measuring villus size, proliferating enterocytes, and brush-border membrane (BBM) enzyme expression and activity. Results showed that thermal conditioning caused an immediate effect characterized by lowered triiodothyronine (T3) level, reduced feed intake, and depressed enterocyte proliferation and BBM enzyme activity. A second series of effects, observed 48 h posttreatment, was characterized by elevated T3, increased feed intake, increased enterocyte proliferation, and higher expression and activity of BBM enzymes. The association between ambient temperature, feed intake, growth rate, and plasma T3 levels was reflected in the structure and function of the intestinal tract. The results suggest that thermal conditioning at an early age influences T3 concentrations, which in turn alter the intestinal capacity to proliferate, grow, and digest nutrients. However, these experiments were not able to discriminate between effects due to feed intake and those due to thermal conditioning. The treatments modulated changes in the intestinal tract following thermal treatment.

Adaptation, Physiological↗

Enterocyte dynamics and mucosal development in the posthatch chick.

Changes in the morphology of the small intestinal mucosa and enterocyte dynamics were examined in posthatch chicks through 12 d. At hatch, enterocytes were round and nonpolar; however, within 24 h posthatch, enterocytes lengthened and exhibited more typical morphology. Crypts were rudimentary at hatch and by 48 h invagination was completed and crypt numbers increased by branching and fission, with the number of crypts per villus reaching plateau after 72 h posthatch. All epithelial cells were proliferative at hatch. In the crypts, the proportion of proliferating enterocytes decreased to 50 to 60% within 2 d posthatch, whereas along the villus the proportion of proliferating cells decreased to 10 to 20% by 6 d. Different patterns of temporal development of villi were observed in the duodenum, jejunum and ileum. Individual villus surface area increased steadily in the duodenum throughout the experiment, whereas individual jejunal and ileal villus surface areas increased more slowly after 4 d posthatch. The number of villi per cross-section of intestine increased in the duodenum and jejunum but not in the ileum. The total segment villus surface area increased similarly in all segments until 3 d posthatch, after which the jejunum increased considerably in absorptive area, whereas the duodenum and ileum increased more slowly. This study shows that, in the hatching chick, the small intestine matures in a manner similar to neonatal mammals, with specific ontogenetic timetables in the different small intestinal segments, however, the most dramatic changes occur within the first 24 h posthatch.

Animals↗

The effect of early feeding on growth and small intestinal development in the posthatch poult.

Poults with early access to feed in the hatchery or turkey house grew more than those reared under standard commercial practice. During 48 h posthatch, fed poults utilized yolk and exogenous feed to increase BW by 11 g. The small intestine increased from 3.8% of BW at hatch to 8.9% after 48 h. In contrast, BW in feed-deprived poults decreased by 10 g, whereas the small intestine increased slightly in weight and composed 4.5% of BW after 48 h. The number of cells per villus and the villus surface area increased dramatically posthatch in the duodenum but more slowly in the jejunum and ileum. Enterocyte width changed little, but length increased more than twofold in the duodenum and by approximately 50% in the jejunum and ileum by 6 d posthatch. Lack of access to feed depressed the rate of growth of villi and enterocyte length in all intestinal segments until 6 d posthatch. All intestinal epithelial cells were proliferating at hatch, which changed rapidly within 48 h posthatch, with proliferating cells becoming located mainly in the intestinal crypts where about half of the cells were proliferating. In feed-deprived poults, the decrease in the proportion of proliferating cells in the crypt was greater than that of fed poults; after refeeding, an increase in the rate of proliferation was observed in feed-deprived poults. Plasma concentrations of Na, glucose, triglycerides, and phospholipids were not affected by feed deprivation; however, nonesterified fatty acid concentrations were enhanced in feed-deprived poults, indicating a greater use of fatty acids for energy. Plasma triiodothyronine (T3) concentrations, which may mediate some of the intestinal effects of feed deprivation, were depressed in poults without access to feed.

Aging↗

Hydrolyzable carbohydrates in pasture, hay, and horse feeds: direct assay and seasonal variation.

Carbohydrates may be hydrolyzed or fermented in the digestive tract, and this distinction is important for the evaluation of the diet of herbivores. Both hydrolyzable and fermentable carbohydrates are included in the nonstructural carbohydrate (NSC) fraction as estimated by difference using proximate analysis. Our objectives were to measure hydrolyzable carbohydrates in forages and concentrates, to compare these values with nonstructural carbohydrate, to test for prediction of hydrolyzable carbohydrate concentration in forages from its near-infrared spectrum, and to examine seasonal variation of carbohydrates in pasture. Samples of forages (107) and concentrates (25) were collected, dried, ground, and analyzed for NSC (calculated as 100 - water - CP - fat - ash - NDF), hydrolyzable carbohydrate (CHO-H, direct analysis), and rapidly fermentable carbohydrate (NSC minus CHO-H). Hydrolyzable carbohydrate accounted for 97% or more of the NSC in the concentrates but only 33% in pasture and hay. A two-term polynomial equation fit all the data: CHO-H = 0.154 x NSC + 0.00136 x NSC2, R2 = 0.98, P < 0.0001, n = 132. In 83 pasture samples, CHO-H concentrations were predicted by near-infrared spectra with a calibration R2 of 0.97, a mean of 48 g/kg, and a SE of calibration of 3.5 g/kg DM. In pasture samples collected between September 1995 and November 1996, the coefficient of variation was 31% for both CHO-H and rapidly fermentable carbohydrate (CHO-FR); the largest increments were 31 g/kg of CHO-H from September to October and 41 g/kg of CHO-FR from February to March. The increased risk of certain diseases, such as laminitis and colic, that have been previously associated with an abrupt overload of NSC may be more precisely attributed to CHO-H in grain concentrates, and to CHO-H as well as CHO-FR in pastures.

Animal Feed↗

Alkanes as internal markers to estimate digestibility of hay or hay plus concentrate diets in horses.

Dry matter intake (DMI), dry matter digestibility (DMD), and fecal output (FO) are difficult to measure directly in the field, and indirect methods using external and internal markers have thus been developed. An experiment was conducted consisting of two digestion trials with two periods in each trial to examine the use of five odd-chain alkanes (C25 to C33) of plant cuticular wax as internal markers to estimate DMD of hay or hay plus concentrate diets in horses. Eight mature Thoroughbred geldings were housed in 4- x 4-m stalls and randomly assigned to one of two mixed grass/legume hays (Diets 1 and 2) in Trial 1 and to mixed grass/legume hay plus one of two concentrates (Diets 3 and 4) in Trial 2. After the first 12-d period was conducted, dietary assignments for each group were switched for the second period in each trial. Each period consisted of a dietary accommodation from d 1 to 7 and total fecal collection from d 8 to 11. Results indicated that fecal recoveries of odd-chain alkanes were 88 to 90% for Diet 1, 75 to 92% for Diet 2, 71 to 81% for Diet 3, and 71 to 82% for Diet 4. Alkane recoveries were not related to alkane chain lengths. Digestibilities calculated from alkane concentration data adjusted using the mean fecal recovery of individual odd-chain alkanes (DA1) were not significantly different from the digestibilities estimated from total collection (DTC) for Diets 1 and 2 in Trial 1 and Diets 3 and 4 in Trial 2. When adjustment was based on the mean recovery of all alkanes (DA2; estimated by linear regression), all DA2 estimates for horses offered all diets were similar to DTC. Results indicate that accurate mean estimates of DMD can be obtained by using plant wax alkane markers and adjusting for the mean recovery of five odd-chain alkanes in a diet.

Alkanes↗

Seasonal changes in bovine fertility: relation to developmental competence of oocytes, membrane properties and fatty acid composition of follicles.

Follicle dynamics and oocyte viability in Holstein primiparous and multiparous cows and the relationships between fertility and the biochemical and physical properties of oocyte membranes with season were examined. The conception rates of primiparous (n = 70 885) and multiparous (n = 143 490) cows differed, peaking in the winter and decreasing in the summer. The number of follicles 3-8 mm in diameter per ovary was higher in winter (19.6) compared with summer (12.0). However, in winter the percentage of ovaries with fewer than ten follicles per ovary was 16%, in contrast to 50% in summer. After aspiration of follicles, 7.5 oocytes per ovary were found in winter and 5.0 oocytes per ovary in summer. Cleavage to the two- to four-cell stage after chemical activation was greater in winter than in summer; this was enhanced at the morula stage and embryo development to the blastocyst stage was significantly higher in winter than in summer. Determination of the lipid phase transition in oocyte membranes revealed a shift of 6 degrees C between summer and winter. Fatty acid composition of phospholipids from follicular fluid, granulosa cells and oocytes indicated that there was a higher percentage of saturated fatty acids during the summer and that the percentages of mono-unsaturated and polyunsaturated fatty acids were higher in oocytes and granulosa cells during the winter. Oocytes and granulosa cells had similar fatty acid compositions, in contrast to follicular fluid. These results may explain the differences in the ability of oocytes to develop to the blastocyst stage at different seasons. Thus, temperature changes may lead to changes in membrane properties, which, in turn, can influence oocyte function and fertility.

Animals↗

Immune responses of chickens to dietary protein antigens. I. Induction of systemic and intestinal immune responses following oral administration of soluble proteins in the absence of adjuvant.

Oral administration of protein antigens in solution leads to the development of oral tolerance in most mammals but rarely so in the chicken. As dietary proteins are not expected to be immunogenic, the present study was undertaken to evaluate immunological consequences following oral exposure to protein antigens in chicks, and to determine whether or not this form of antigen is ignored. Chicks and turkey poults were fed solutions containing bovine serum albumin (BSA), porcine serum albumin, beta-lactoglobulin or bovine hemoglobin over a period of 6 days (25mg/chick/day). At different time points after feeding serum and bile were examined for presence of specific antibodies by ELISA. Surprisingly, the fed antigens induced robust antibody responses in the absence of added adjuvant. This immune response was further characterised to show that (1) a daily feeding regimen was more immunogenic than single dose feedings, (2) by using a daily feeding regimen, as little as 2mg/chick/day was fully immunogenic, (3) effective immunization was attained in chicks older than 10 day of age, (4) the main antibody class in the serum was IgG, and (5) high IgA levels were detected in the bile after booster feedings. These observations are difficult to reconcile with current concepts on peripheral tolerance to innocuous antigens, and indicate that the bird regulates tolerance and response in a manner different from that described in mammals.

Adjuvants, Immunologic↗

Linseed oil suppresses the anti-beta-2-glycoprotein-I in experimental antiphospholipid syndrome.

The immunomodulatory potential of a diet enriched with n-3 polyunsaturated fatty acids was analysed in naïve mice with experimental antiphospholipid syndrome (APS) induced by active immunization with H-3, a human anti-beta-2-glycoprotein-I (anti-betaGPI) monoclonal antibody (mAb). Fetal loss and other clinical manifestations of APS were prevented in the group of APS mice upon exposure to the enriched n-3 diet compared to the control group. The titers of anti-betaGPI were significantly lower (in O.D. at 405 nm, 1.387+/-0.232 in comparison to non-treated mice 0.637+/-0. 111, P< 0.05). The reduced titer of anti-betaGPI antibodies in the sera of the treated mice was associated with a reduced number of anti-betaGPI forming cells in cultured splenocytes (84+/-14, antibody-forming cells (AFC)/10(5)cells in comparison to 37+/-4 AFC/10(5)cells (P< 0.02).In addition to the suppression of the humoral response in mice with experimental APS fed with linseed oil, we also observed an inhibitory effect on the cellular response. The T-cell response to anti-betaGPI was lower in comparison to mice immunized with H-3 anti-betaGPI mAB, which were kept on a normal diet. These results indicate that polyunsaturated fatty acids may improve clinical and laboratory parameters of APS. The beneficial effects of diets enriched with n-3 should be further examined as a potential mode of therapy for patients with APS.

Animals↗

Small intestinal development in the young chick: crypt formation and enterocyte proliferation and migration.

1. Post-natch mucosal development was examined in the chick small intestinal epithelium using immunostaining with proliferating cell nuclear antigen (PCNA) and 5-bromo-2-deoxyuridine (BrdU). 2. On the day of hatching jejunal crypts were small and a single crypt per villus was observed. However, during the 108 h post-hatch crypts developed rapidly branching and increasing in size, cell numbers and cell size. 3. Almost all epithelial cells in the small intestine of the hatching chick were proliferating, as indicated by PCNA and BrdU, while more than 80% of proliferating cells were localised in the crypts after 108 h post hatch. 4. Estimate of villus cell transit time using BrdU was only possible from 48 h post-hatch when villus transit time was 72 h in the jejunum, whereas at 336 h transit time was 96 h. 5. In the 108 h post hatch a rapid transition occurs from total jejunal epithelial cell proliferation and immature crypts to a defined proliferative zone in the crypts, with constant division and migration.

Animals↗

Effect of age of turkeys on the metabolisable energy of different foodstuffs.

1. True metabolisable energy (TME) of various foodstuffs were determined in turkeys at 3, 9 and 17 weeks of age. 2. TME of soyabean meal did not change with age but TME of wheat and maize was 13% higher in 17-week-old turkeys than in 3- and 9-week-old birds. 3. These differences should be taken into account when formulating diets for turkeys.

Aging↗

Early posthatch starvation decreases satellite cell proliferation and skeletal muscle growth in chicks.

The effect of posthatch starvation on skeletal muscle growth and satellite cell proliferation was examined in chicks. Chicks were either fed or starved for 48 h posthatch (d 0-d 2, d 2-d 4 or d 4-d 6) and then refed for 41 d. Body and breast muscle weights were significantly lower in starved chicks than in fed controls throughout the experiment. Histochemical staining revealed that skeletal muscle fiber development in the starved group lagged behind that of the fed group. Starvation from d 2 to 4 and d 4 to 6 posthatch had a progressively lesser effect than did immediate posthatch starvation (P < 0.05). In vitro culturing of breast muscle satellite cells revealed that DNA synthesis and number of cells per gram of muscle in the fed chicks peaked on d 2 and d 3, and then declined. In contrast, DNA synthesis in the cells of starved chicks declined on d 2 and increased on d 3 when chicks were refed. A similar pattern was seen for the number of cells per gram muscle; however, in general cell numbers tended to be higher in the starved group than in controls (P < 0.1). The results obtained with cultured cells were parallel with in situ immunostaining with 5-bromo-2'-deoxyuridine and proliferating cell nuclear antigen in breast muscle from experimental chicks, and with growth hormone receptor expression. These results suggest that satellite cell cultures are a reliable tool for evaluating muscle growth in postnatal chickens. We conclude that sufficient feed in the immediate postnatal period is critical for satellite cell proliferation and skeletal muscle development and is thus important for optimal muscle growth.

Aging↗

Partial sequence and expression of the gene for and activity of the sodium glucose transporter in the small intestine of fed, starved and refed chickens.

A 970 bp cDNA Na(+)/glucose cotransporter (SGLT1) was isolated and sequenced from chicken jejunum by reverse transcriptase polymerase chain reaction (RT-PCR) using primers based on conserved regions. Using the 970 bp PCR product as a specific probe, Northern Blot hybridization indicated a transcript of ca. 4 kb. The isolated chicken intestinal SGLT1 cDNA was used to quantitate mRNA expression. Glucose uptake activity and kinetics were determined in brush border membrane vesicles (BBMV) from jejunum tissue of chickens which were either fed, food-deprived or refed following food deprivation. Net glucose uptake to BBMV was higher (P: < 0.02) in the control and refed chicks (149 +/- 11.9, 139.6 +/- 7.43 pmol x mg protein(-1) x s(-1)) than in food-deprived chicks (107 +/- 4.23 pmol x mg protein(-1) x s(-1)). The k(m) (150 micromol/L) and V:max (1111.1 pmol x mg protein(-1) x s(-1)) were higher in the food-deprived chicks compared to control and refed birds (25, 24 micromol/L and 227,142 pmol x mg protein(-1) x s(-1), respectively). Expression of SGLT1 mRNA was significantly enhanced in the food-deprived and refed birds. In food-deprived chicks the lower affinity and higher activity of the SGLT1 transporter for glucose were accompanied by higher expression of mRNA which might indicate that the transporter was upregulated by low substrate concentration. Quantification of expression of intestinal mRNA of SGLT1 provides important information concerning control of nutrient uptake.

Amino Acid Sequence↗

Hydrolysis and absorption in the small intestines of posthatch chicks.

In the immediate posthatch period, chicks must transfer from metabolic dependence on yolk to utilization of exogenous feed. This study describes changes in intestinal luminal pancreatic enzyme activity and mucosal uptake posthatch as influenced by feed and Na intake. Chicks with access to feed increased in BW and small intestinal weight in the 48-h posthatch, whereas chicks without access to feed decreased in BW; however, small intestinal weight increased during this period. Chicks ingesting feed showed increases in total intestinal trypsin, amylase and lipase activities that were correlated with intestinal weights and BW. Chicks without access to feed showed little change in trypsin and amylase activities, and these increased only after feed consumption. Feeding a low-Na diet did not significantly change the regression coefficient between pancreatic enzyme activity and BW. Mucosal uptake was estimated by measuring Na+,K+-adenosine triphosphatase (ATPase) activity in small intestinal segments. In fed birds this activity increased in relationship to growth, whereas in nonfed birds uptake increased only after access to feed. Low-Na diets allowed only minimal mucosal uptake in all intestinal segments. This study indicates that secretion of trypsin and amylase into the intestine was triggered by feed intake. In addition, Na plays a critical role in intestinal uptake in the immediate posthatch period.

Amylases↗