Influence of inclusion level of barley in wheat-based diets and supplementation of carbohydrase on growth performance, nutrient utilisation and gut morphometry in broiler starters.

1. The influence of barley inclusion level and supplementation of a multi-component non-starch polysaccharide degrading enzyme on performance and nutrient utilisation in broilers was investigated. Normal-starch hulled barley was evaluated with five levels of inclusion (0, 141, 283, 424 and 565 g/kg) in a wheat-based diet and two levels of enzyme supplementation (0 and 150 g/tonne of feed; a 5 × 2 factorial arrangement of 10 dietary treatments). All diets were equivalent in metabolisable energy and digestible amino acid contents. A total of 400, one-d old male broilers (five cages/treatment; eight birds/cage) were used in the experiment.2. Regardless of enzyme supplementation, weight gain (WG) increased up to 283 g/kg of barley and was reduced afterwards (P < 0.01). Increasing levels of barley resulted in greater (P < 0.001) gain per feed (G/F). Enzyme addition increased WG (P < 0.05) and G/F (P < 0.001) at each barley inclusion level.3. Birds fed diets with 0 and 565 g/kg barley showed the lowest and highest (P < 0.001to 0.05) digestibility for all nutrients measured, respectively. Digestibility of all nutrients was improved by enzyme supplementation at each barley inclusion level (P < 0.05). The nitrogen-corrected apparent metabolisable energy improved with increasing inclusion of barley (P < 0.001) and supplemental enzyme (P < 0.01). Increasing inclusion of barley increased the relative weight of gizzard (P < 0.001) and reduced jejunal digesta viscosity (P < 0.001). Supplemental enzyme (P < 0.001) reduced digesta viscosity.4. The optimum inclusion level of barley, with respect to growth performance, was 283 g/kg of diet. Increasing barley inclusion improved nutrient and energy utilisation, possibly through lowered digesta viscosity and better function of the gizzard. Feed efficiency and nutrient and energy utilisation can benefit from carbohydrase supplementation in barley-based diets, regardless of barley inclusion level.

Developmental morphology study on the stomach of African ostrich chicks.

This study investigates the developmental morphology changes on the proventriculus and gizzard of African ostrich chicks using gross anatomy, and light and transmission electronic microscopy. The body weight; the weight of the proventriculus and gizzard; the thickness of simple tubular glands, muscularis mucosae, compound tubular glands, and muscular layer of the proventriculus; and the thickness of the gizzard glands and muscularis mucosae were measured on postnatal d one, 45, 90, and 334. Under transmission electronic microscopy, the oxynticopeptic cells, mucous cells, and endocrine cells were observed in the proventriculus on postnatal d one, 45, and 90. Our results revealed that the weights of the proventriculus and gizzard (relative to the body weight) both peaked on d 90, respectively, and declined thereafter. The thicknesses of the simple tubular glands, muscularis mucosae, compound tubular glands, muscular layer of the proventriculus, and gizzard glands increased with increasing bird age. However, the thickness of the muscularis mucosae in the gizzard peaked on d 90 and gradually decreased thereafter. The result of transmission electron microscopy revealed that the glands in the submucosa layer of the proventriculus did not differ from other cell types, aside from an increase in cell diameter and an increase in cytoplasmic content. A significant increase in the number of mitochondria and smooth endoplasmic reticulum was noted, as well as an in granules secreted by endocrine cells. Therefore, to improve brood rates of reared African ostrich chicks, feed management ought to be enhanced between postnatal d one and day 90.

Effects of dietary coarsely ground corn and 3 bedding floor types on broiler live performance, litter characteristics, gizzard and proventriculus weight, and nutrient digestibility.

The effects of zero or 50% dietary coarsely ground corn (CC) in pelleted and screened grower and finisher diets on broilers reared on 3 bedding floor types (plastic net [NET], new pine wood shavings litter [NEW], or old pine wood shavings litter [OLD]) on broiler live performance, litter characteristics, gizzard and proventriculus weight, and apparent ileal digestibility (AID) were studied in a 2 × 3 factorial arrangement of treatments. Fine corn was produced with a hammermill (271 µm) and CC with a roller mill (1145 µm). Utilization of CC reduced milling cost by 9.47 cents per MT with similar nutrient content of screened pellets. The 50% CC treatment exhibited improved (P ≤ 0.05) feed intake at 42 d (2.5%) and 49 d (3.0%), and BW (5.4%) and FCR from 28 d (1.4%). Birds on NEW litter exhibited improved (P < 0.05) BW at 28 and 35 d and 42 d FCR as compared to NET and 49 d FCR compared to NET and OLD. The 50% CC treatment exhibited increased (P < 0.05) gizzard weight but decreased proventriculus weight at 49 d. NEW litter birds exhibited increased (P ≤ 0.05) gizzard weight at 28 d and 49 d and decreased proventriculus weight as compared to NET at 49 days. The 50% CC treatment exhibited decreased (P ≤ 0.05) litter moisture at 35 and 42 d, litter N at 35 and 49 d, and litter pH at 49 days. OLD litter birds exhibited greater (P < 0.05) litter N at 14, 35, and 49 d, as well as litter moisture, pH, and ammonia concentration at 49 days. The 50% CC group also exhibited improved AID of nitrogen (P < 0.05). Broilers fed pelleted and screened diets containing 50% CC exhibited improved live performance and reduced litter moisture while use of NEW litter resulted in a somewhat similar effect, which indicated that consumption of NEW litter also facilitated gastric development and function.

Correlation of ghrelin concentration and ghrelin, ghrelin-O-acetyltransferase (GOAT) and growth hormone secretagogue receptor 1a mRNAs expression in the proventriculus and brain of the growing chicken.

To determine mechanisms for age-related decrease of GHS-R1a expression in the chicken proventriculus, changes in mRNA expression of ghrelin and ghrelin-O-acetyltransferase (GOAT) as well as ghrelin concentrations in the proventriculus and plasma were examined in growing chickens. Changes in expression levels of ghrelin, GOAT and GHS-R1a mRNAs were also examined in different brain regions (pituitary, hypothalamus, thalamus, cerebellum, cerebral cortex, olfactory bulb, midbrain and medulla oblongata). Ghrelin concentrations in the proventriculus and plasma increased with aging and reached plateaus at 30-50 days after hatching. High level of ghrelin mRNA decreased at 3 days after hatching, and it became stable at half of the initial level. Expression levels of GHS-R1a and GOAT decreased 3 or 5 days after hatching and became stable at low levels. Significant negative correlations were found between plasma ghrelin and mRNA levels of GOAT and GHS-R1a. Expression levels of ghrelin mRNA were different in the brain regions, but a significant change was not seen with aging. GHS-R1a expression was detected in all brain regions, and age-dependent changes were observed in the pituitary and cerebellum. Different from the proventriculus, the expression of GOAT in the brain increased or did not change with aging. These results suggest that decreased GHS-R1a and GOAT mRNA expression in the proventriculus is due to endogenous ghrelin-induced down-regulation. Expression levels of ghrelin, GOAT and GHS-R1a in the brain were independently regulated from that in the proventriculus, and age-related and region-dependent regulation pattern suggests a local effect of ghrelin system in chicken brain.

Improving the efficiency of feed utilization in poultry by selection. 1. Genetic parameters of anatomy of the gastro-intestinal tract and digestive efficiency.

BACKGROUND: Feed costs represent about 70% of the costs of raising broilers. The main way to decrease these costs is to improve feed efficiency by modification of diet formulation, but one other possibility would be to use genetic selection. Understanding the genetic architecture of the gastro-intestinal tract (GIT) and the impact of the selection criterion on the GIT would be of particular interest. We therefore studied the genetic parameters of AMEn (Apparent metabolisable energy corrected for zero nitrogen balance), feed efficiency, and GIT traits in chickens.Genetic parameters were estimated for 630 broiler chickens of the eighth generation of a divergent selection experiment on AMEn. Birds were reared until 23 d of age and fed a wheat-based diet. The traits measured were body weight (BW), feed conversion ratio (FCR), AMEn, weights of crop, liver, gizzard and proventriculus, and weight, length and density of the duodenum, jejunum and ileum. RESULTS: The heritability estimates of BW, FCR and AMEn were moderate. The heritability estimates were higher for the GIT characteristics except for the weights of the proventriculus and liver. Gizzard weight was negatively correlated with density (weight to length ratio) of duodenum, jejunum and ileum. Proventriculus and gizzard weights were more strongly correlated with AMEn than with FCR, which was not the case for intestine weight and density. CONCLUSIONS: GIT traits were largely dependent on genetics and that selecting on AMEn or FCR would modify them. Phenotypic observations carried out in the divergent lines selected on AMEn were consistent with estimated genetic correlations between AMEn and GIT traits.

Ontogeny of ghrelin mRNA expression and identification of ghrelin-immunopositive cells in the gastrointestinal tract of the Peking duck, Anas platyrhynchos.

Ghrelin is an acylated peptide and an endogenous ligand for the growth hormone secretagogue receptor (GHS-R), and stimulates growth hormone release and food intake in mammals. Peking duck is a very fast growing species of poultry. Although the sequence and structure of ghrelin have recently been determined, the expression of ghrelin in Peking duck has not been studied. Here, we investigated the tissue expression and distribution of ghrelin by RT-PCR and immunohistochemistry, respectively, in Peking duck at different stages of development. Ghrelin mRNA expression was mainly detected in the proventriculus and proventriculus-gizzard junction. It was first expressed, but weakly, on embryonic day 14 (E14); the expression increased by embryonic day 21 (E21), and was maintained at high levels between post-hatching-day 1 (P1) and post-hatching-day 60 (P60). Weak expression of ghrelin mRNA was also found in the gizzard and duodenum. In the gastrointestinal tract of growing Peking duck in P60, the largest number of ghrelin-ip cells was detected in the epithelium of the compound tubular glands in the proventriculus and the next largest number was in the proventriculus-gizzard junction. Very few ghrelin-ip cells were located in the epithelium of the simple tubular glands adjacent to the gizzard. No ghrelin-ip cells were observed elsewhere in the gastrointestinal tract. Ghrelin-ip cells were found in embryos as early as day E21; at the same time, the compound tubular glands in the proventriculus had formed. The numbers of ghrelin-ip cells on P1 were similar to those of E21 embryos. However, on P60, high numbers of strongly stained ghrelin-ip cells were found to be scattered in the epithelium of the compound tubular glands in the proventriculus. The density of ghrelin-ip cells (cells/mm(2)) in the proventriculus on P60 was significantly greater than those of P1 and E21 embryos. These results demonstrate that ghrelin is expressed in the Peking duck gastrointestinal tract, especially in the proventriculus, from mid-late-stage embryos to growing period and suggested an involvement of ghrelin in the development and biology of the gastrointestinal tract of the Peking duck.

Distribution and developmental change of lymphoid tissues in the chicken proventriculus.

In the chicken proventricular mucosa, aggregations of lymphocytes were localized in three different sites of the lamina propria, namely, underneath the surface epithelium, near the duct orifice of the deep proventricular gland, and in the gland tissue itself. In the lymphoid masses underneath the surface epithelium and in those near the duct orifice, CD4+ T lymphocytes and TCR2+ T lymphocytes occupied their central part, and B lymphocytes were localized in the periphery. CD8+ T lymphocytes and TCR1+ lymphocytes were evenly distributed in the masses. Infiltration of lymphocytes into these sites was first observed on the 20th embryonic day. At 1 week after hatching, CD3+ lymphocytes began to occupy the central area of the masses and His-C1+ B lymphocytes tended to be located in the periphery. Ultrastructurally, M cells were found neither in the epithelium of the mucosa nor in that of the excretory duct close to the lymphoid masses. In the deep proventricular gland, the lymphoid masses had a germinal center consisting of B lymphocytes, surrounded by the T lymphocyte-rich periphery. These masses were first recognized at the 3rd post-hatching week, presumably being formed against possible antigens invading into the lumen of the proventricular gland. On the other hand, the lymphoid masses beneath the surface epithelium and those near the duct orifice existing before the hatching period were considered to be prepared to establish the local mucosal immune barriers against the expectant antigenic invasion.

Growth of the digestive organs in ducks with considerations on their growth in birds in general.

1. Growth of the oesophagus, proventriculus, gizzard, intestine, liver and pancreas weight was investigated in Mallards, White Pekins, Muscovies and a Muscovy x White Pekin cross. The birds varied in age between hatching and 154 d. The data were analysed by fitting both the Janoschek growth curve and the allometric formula. 2. The growth rate of all organs, except the oesophagus, peaked earlier (30 d for Muscovies and 14 d for the other breeds) than body weight and they grew faster to any given percentage of their final weight. In contrast, oesophagus weight showed growth curve characteristics similar to body weight. 3. Oesophagus weight showed simple, slightly negative allometry. The remaining organs followed complex allometry that can approximately be described by 2 allometric stages. The 1st phase was isometric to positive allometric. The 2nd showed marked negative allometry. 4. These growth patterns are assumed to be generally present in birds.

Performance and gastro-intestinal response of broiler chickens fed on cereal grain-based foods soaked in water.

1. Two experiments were carried out to investigate the addition of 1 3 kg water per kg air-dry mash diets containing high proportions (600 to 700 g/kg) of ground cereal grains (wheat, barley or oats) on broiler performance and the structure and function of the gastro-intestinal tract. 2. Chicks at the age of 7 d were fed on the wheat-, barley- or oats-based diets in the dry or wet forms for 35 d. Food and water intakes were recorded daily while body weight was measured weekly. Two birds from each treatment were killed each week to measure gut size and the viscosity of gut contents. Tissue samples from various digestive segments were histo-morphologically examined to determine the thickness of tissue layers, size of tissue glands, villa heights, crypt depths and thickness of tunica muscularis. Crypt cell proliferation rate (CCPR) for each segment was also determined using a metaphase arrest technique. 3. The results from both experiments showed that wetting food significantly (P<0.05) increased food intake, total water intake and body weight gain of broiler chickens. The body weight gains of birds were proportional to their food intakes so that the efficiency of food utilisation was similar for all treatments. Dry matter retention of food tended to increase in birds given wet food from 7 to 21 d but not thereafter, compared to the dry-fed birds. Although water intake from the water bottle was significantly (P<0.05) reduced in birds given wet food, total water intakes from the water bottle plus that from food were significantly (P<0.05) higher in the wet-fed birds than in the dry-fed birds. The ratio of total water to dry food intake was, however, similar in both feeding regimens. 4. The fresh empty weight of the gut was increased by wet-feeding while its relative weight to body weight and the length of gut was not affected by dietary treatments. Significantly greater development of the tissue glands in the proventriculus and gizzard was observed in the birds given wet food; this was associated with the reduced thickness of the muscular layer of these segments. An increase in villus height was also observed in duodenum, small intestine, caeca and colon of birds given wet food, compared to those given dry food. CCPR was significantly (P<0.05) reduced by wet-feeding throughout the digestive tract. This was associated with a significant decrease in the mean viscosity of the gut contents and the concentration of volatile fatty acids (VFAs) in the caeca. 5. Wetting diets based on cereal grains caused a significant improvement in the performance of broiler chickens. The mechanism of the beneficial effects of wet feeding could be attributed to the decreased viscosity of gut contents; the greater development of the layer of villi in the digestive segments and the reduced CCPR in the crypts of the epithelium.

Development of the diffuse endocrine system in the chicken proventriculus.

The development of endocrine cells in the chicken proventriculus has been investigated using light- and electron-microscopy in conjunction with silver and immunocytochemical techniques. The first morphologically detectable endocrine cells were found in 5-day-old embryos by electron microscopy. From the 9th to the 13th day, endocrine cells in contact with the lumen of the organ could be detected both by electron and light (silver impregnation) microscopy. The number of open-type endocrine cells progressively decreased and the number of closed-type increased after this stage. Until the 16th day, endocrine cells were located exclusively in the luminal epithelium, but afterwards they appeared in progressively greater numbers in the compound glands. After hatching, long cytoplasmic processes could be seen in the endocrine cells. Immunoreactivities to regulatory substances appeared in the following order: serotonin (day-14), avian pancreatic polypeptide, glucagon and somatostatin (day-16), bombesin and neurotensin (day-18), and finally, met-enkephalin (day-21).

Development and birthdates of vasoactive intestinal peptide immunoreactive neurons in the chick proventriculus.

To gain insight into the mechanisms regulating expression of transmitter phenotypes in the enteric nervous system, we have studied the development and birthdate of vasoactive intestinal peptide immunoreactive (VIP-IR) myenteric neurons in the chicken proventriculus (secretory portion of the avian stomach) by a combination of immunocytochemistry and radioautography. The appearance and numbers of VIP-IR neurons in whole mounts of the myenteric plexus from chick embryos and chickens were examined. We found that VIP-IR neurons first appeared at embryonic day (E) 5.5-6.5 in the distal part of the proventriculus. At E7.5, VIP-IR neurons were found singly, in pairs, or in small groups, which together with unlabeled cells formed primitive myenteric ganglia. VIP-IR fibers were found within the developing fiber tracts which connected the ganglia. The number of VIP-IR neurons was found to be maximum in the E15.5 embryo and to decline to 68% of maximum in the 4 week old chicken. Birthdate studies were performed by application of either single pulses or cumulative doses of [3H]-thymidine to embryos between E3 and E14. Whole mounts of the myenteric plexus from the proventriculus of these embryos were immunostained for VIP at E10 or E17. The whole mounts were subsequently sectioned and processed for radioautography. We found that VIP-IR myenteric neurons were born between E3 and E10 with a peak at E7. Most cells underwent terminal division between E5 and E9. These data will be useful in determining the time and conditions when cells make decisions about transmitter phenotypes.

Organ growth and digestive enzyme levels to fifteen days of age in lines of chickens differing in body weight.

Weights of internal organs and levels of digestive enzymes were obtained through the first 15 days posthatch for cockerels from three lines of chickens known to differ greatly in body weight. On Day 15 body weights from the fastest growing line were eight times greater than those from the slowest growing line. Differences among lines were found for weights at hatching and for growth patterns (both absolute and relative to body weight) of the vitelline residue, heart, lungs, liver, pancreas, crop, proventriculus, gizzard, and segments of the small intestine. Line differences were also evident for levels of trypsin, chymotrypsin, and amylase in the pancreas and contents of the small intestine. Ranking of lines for these traits varied with age. In all lines weights of the small intestine, liver, and pancreas increased relatively more than did total body weight during the 1st wk posthatch, after which the relationship reversed.

Formation of ganglia in the gut of the chick embryo.

We have examined the formation of myenteric ganglia in the developing avian enteric nervous system. The monoclonal antibody HNK-1 was used to identify neural-crest-derived cells in whole mounts of fore- and midgut of chick embryos. We find that the crest-derived cells extend processes to their neighbors and form a complex network in the wall of the gut. Formation of this network is an unusual behavior of crest-derived cells and suggests the gut microenvironment is critical to this behavior. This cellular network disappears after ablation of the vagal neural crest, indicating the HNK-1-stained cellular network arises from crest-derived cells. The network is found in the gut wall before the vagal nerve fibers are present. This network is first found in the primordium of the proventriculus, distal to the evagination of the lung buds, and progresses just proximal to the yolk stalk at embryonic day (E) 3.5 and almost to the ileocecal junction at E5.5. The number of cells and the complexity of the network decrease in a rostral-caudal direction down the length of the gut at these stages. The leading edge of the network consists of cells serially arranged in longitudinally running strands. The organization of the network changes with increasing embryonic age; we have focused on network changes in the proventriculus. In the primordium of the proventriculus at E3.5, the network consists of a cluster of one or two adjacent crest-derived cells, which extend processes to a number of neighboring crest-derived cells. At E5.5 large increases in the number of cells per cluster and in the length of cellular connectives between clusters are apparent. At E6.5 a crude meshwork of clusters is seen. At E10.5 the arrangement of cell clusters resembles the pattern of ganglia found in the adult myenteric plexus. This network may provide the environmental cues for the differentiation of enteric neurons and a framework for the pattern of ganglia found in the adult enteric nervous system.

Developmental patterns of selected characteristics of the gastrointestinal tract of young turkeys.

Developing embryos and hatchling poults were sampled (n = 4) at Days 22, 24, 26, and 28 of incubation and at 1, 2, 4, 6, and 8 days after hatching, and selected characteristics of the gastrointestinal tract (GIT) were measured. Body weight increased linearly up to day of hatching and also from 2 to 8 days posthatching. Residual yolk weight decreased rapidly starting on Day 26 of incubation and was nearly depleted by 4 days posthatching. Changes in weight of segments of the GIT nearly paralleled the increase in body weight until day of hatching. Thereafter, weights of the proventriculus, small intestine, and pancreas increased more rapidly than body weight until 6 days after hatching. At this time, change in weight of small intestine and pancreas seemed to parallel that of body weight, whereas proventriculus weight continued to increase more rapidly. Gizzard weight, as a percentage of body weight, increased until Day 4 posthatching and then remained relatively constant through 8 days. Specific activities (SA) of pancreatic amylase, lipase, and trypsin were low until after hatching. Subsequently, amylase SA increased nearly threefold by Day 6. Lipase SA remained nearly constant between Days 1 and 8, and trypsin SA increased only slightly. Total activities of pancreatic enzymes, however, increased substantially after hatching, mainly because of increased pancreas weight. Jejunal maltase SA was high at hatching but decreased markedly by Day 4. This decrease in SA resulted in a notable reduction in total maltase activity of the jejunum despite an increase in jejunum weight.

Postnatal development of mucosa-associated lymphoid tissues in chickens.

The postnatal development of chicken mucosa-associated lymphoid tissues of the eyes, lungs, and intestines were investigated with monoclonal antibodies specific for either all leucocytes, B lymphocytes, mononuclear phagocytes, IgM, IgG, or IgA. Attention has been paid to the relation of lymphoid infiltrates with their surrounding mucosae, the segregation into B-cell and T-cell areas, development of germinal centers, and secretory immunoglobulins. Abundant secretory IgM and IgA was detected in the epithelium of the Harderian glands in the orbits, even though they lacked large leucocyte infiltrates with germinal centers. Lymphoid tissues in the mucosae of lungs and intestines developed separate B-cell and T-cell areas. The proventriculus, Meckel's diverticulum, and Peyer's patches generally contained germinal centers from 12 weeks of age on. Because chickens as young as 2 weeks old had germinal centers in bronchus-associated lymphoid tissue and cecal tonsils, these areas were probably highly stimulated by antigens. Isotype-specific monoclonal antibodies were used to detect IgM-, IgG-, and IgA-bearing follicular cells in the same germinal center.

Ontogenesis of bombesin-like immunoreactive cells in the chicken proventriculus.

The distribution and morphological behaviour of bombesin like immunoreactive cells in the proventriculus of chick embryos, newborns and adults were investigated by indirect immunoperoxidase procedure. The first immunoreactive cells appear around the 11th day of incubation. Initially few, they increase progressively to reach a peak around the 18th day. Throughout the embryonic period almost all immunoreactive cells are column-shaped, display a typical "open" behaviour, and are situated among other superficial lining epithelial cells. Around hatching time, these "open" cells suddenly decrease in number while another immunoreactive "closed" cell type becomes more evident in a deeper location. In newborns and adults, the majority of immunoreactive cells are oval shaped and are characteristically situated near the neck of lobular glands; they seem to display a "closed" behaviour. The results demonstrate two different kinds of peptide-containing cells which probably exert different functions. During the embryonic period, in fact, they resemble a chemioreceptor, but in postnatal life they appear as a paracrine/endocrine elements.