Influence of method of whole wheat inclusion and xylanase supplementation on the performance, apparent metabolisable energy, digestive tract measurements and gut morphology of broilers.

1. The aim of the present study was to examine the influence of whole wheat inclusion and xylanase supplementation on the performance, apparent metabolisable energy (AME), digesta viscosity, and digestive tract measurements of broilers fed on wheat-based diets. The influence of the method of whole wheat inclusion (pre- or post-pelleting) was also compared. A 3 x 2 factorial arrangement of treatments was used with three diet forms (648 g/kg ground wheat [GW], GW replaced by 200 g/kg of whole wheat before [WW1] or after cold-pelleting [WW2]) and two xylanase levels (0 and 1000 XU/kg diet). 2. Birds given diets containing whole wheat had improved weight gains, feed efficiency and AME compared to those fed on diets containing ground wheat. The relative gizzard weight of birds fed WW2 diets was higher than in those fed GW and WW1 diets. Pre-pelleting inclusion of whole wheat had no effect on relative gizzard weight. Post-pelleting inclusion of whole wheat resulted in greater improvements in feed efficiency and AME than the pre-pelleting treatment. 3. Xylanase supplementation significantly improved weight gain, feed efficiency and AME, irrespective of the wheat form used. Viscosity of the digesta in the duodenum, jejunum and ileum were reduced by xylanase addition. Xylanase supplementation reduced the relative weight of the pancreas. 4. Neither xylanase supplementation nor whole wheat inclusion influenced the relative weight and length of the small intestine. 5. Xylanase supplementation increased ileal villus height. A significant interaction between diet form and xylanase was observed for ileal crypt depth. Xylanase supplementation had no effect on crypt depth in birds fed on diets containing GW, but increased the crypt depth in WW2 diets. No significant effects of diet form and xylanase supplementation were observed for the thickness of the tunica muscularis layer of gizzard or villus height, crypt depth, goblet cell numbers or epithelial thickness in the ileum. 6. Improved performance observed with post-pelleting inclusion of whole wheat was associated with increased size of the gizzard and improved AME. The gizzard development hypothesis, however, will not explain the improvements observed with pre-pelleting inclusion of whole wheat, suggesting the involvement of other factors.

Influence of phytase and xylanase, individually or in combination, on performance, apparent metabolisable energy, digestive tract measurements and gut morphology in broilers fed wheat-based diets containing adequate level of phosphorus.

1. The aim of the present study was to examine the influence of microbial phytase and xylanase, individually or in combination, on performance, apparent metabolisable energy, digesta viscosity, digestive tract measurements and gut morphology in broilers fed on wheat-soy diets containing adequate phosphorus (P). The wheat-soy basal diet was formulated to contain 4.5 g/kg non-phytate P and the experimental diets were formulated by supplementing the basal diet with xylanase (1000 xylanase units/kg diet), phytase (500 phytase units/kg diet) or a combination of phytase and xylanase. 2. Supplemental phytase improved the weight gains and feed efficiency by 17.5 and 2.9%, respectively. Corresponding improvements due to the addition of xylanase were 16.5 and 4.9%, respectively. The combination of phytase and xylanase caused no further improvements in broiler performance. 3. Individual additions of xylanase or phytase resulted in numerical improvements in apparent metabolisable energy (AME), but the differences were not significant. The combination of the two enzymes significantly increased AME. Addition of xylanase and the combination of the two enzymes reduced the viscosity of digesta in all sections of the intestine. Phytase supplementation reduced digesta viscosity in the duodenum and ileum, but not in the jejunum. 4. Enzyme supplementation lowered the relative weight and length of the small intestine. Additions of xylanase and phytase reduced the relative weight of the small intestine by 15.5 and 11.4%, respectively, while the corresponding reductions in the relative length of the small intestine were 16.5 and 14.1%, respectively. The combination of phytase and xylanase had no further effects on the relative weight and length of the small intestine compared with the xylanase group. 5. The addition of phytase increased villus height in the duodenum and decreased the number of goblet cells in the jejunum compared with those on the unsupplemented basal diet. Xylanase supplementation tended to increase goblet cell numbers in the duodenum and decreased crypt depth in thejejunum. The combination of phytase and xylanase increased villus height in the ileum and crypt depth in thejejunum and ileum. 6. In summary, the present results showed that the addition of a microbial phytase, produced by solid state fermentation and containing significant activities of beta-glucanase and xylanase, was as effective as xylanase in improving the performance of broiler chickens fed on wheat-based diets containing adequate levels of P. Improved performance with enzyme supplementation was generally associated with reduced digesta viscosity, increased AME, and reduced relative weight and length of small intestine.

Immunohistochemical detection of myogenic cells in muscles of fetal and neonatal lambs.

Previous studies have implied that myonuclei accumulation in a muscle is more important than myofibre number in the determination of muscle size in fetal/neonatal lambs. However, due to the lack of a reliable marker, the role of myogenic precursor nuclei (satellite cells) in myofibre hypertrophy in late fetal and postnatal life is not well understood. In this study, MyoD was shown to be a useful marker for actively proliferating satellite cells in both fetal and neonatal lambs. MyoD was used to determine whether there were differences in the number of actively proliferating satellite cells between single and twin fetuses/neonates, which may explain at least some of the difference in myofibre size observed near birth. Eighteen single-bearing and 9 twin-bearing Coopworth ewes were randomly assigned to one of three slaughter groups (100, 120 and 140 days of gestation). The remaining ewes were kept on pasture until 20 days postpartum at which time 4 single and 4 twin lambs were sacrificed. Twin fetuses/neonates had lower body weights and muscle weights compared to singles. Lower muscle weights in the twins were associated with smaller myofibre cross-sectional areas and lower total nuclei numbers and myogenic precursor cell numbers per muscle in selected hind-limb muscles. These results indicate that myofibre hypertrophy in late gestation and early postnatal life is related to myogenic precursor cell number which may have important implications for growth potential of the growth-restricted fetus.

Safety assessment of potential probiotic lactic acid bacterial strains Lactobacillus rhamnosus HN001, Lb. acidophilus HN017, and Bifidobacterium lactis HN019 in BALB/c mice.

The general safety of immune-enhancing lactic acid bacteria (LAB) strains Lactobacillus rhamnosus HN001 (DR20), Lb. acidophilus HN017, and Bifidobacterium lactis HN019 (DR10) was investigated in a feeding trial. Groups of BALB/c mice were orally administered test LAB strains or the commercial reference strain Lb. acidophilus LA-1 at 2.5 x 10(9), 5 x 10(10) or 2.5 x 10(12) colony forming units (CFU)/kg body weight/day for 4 weeks. Throughout this time, their feed intake, water intake, and live body weight were monitored. At the end of the 4 week observation period, samples of blood, liver, spleen, kidney, mesenteric lymph nodes, and gut tissues (ileum, caecum, and colon) were collected to determine: haematological parameters (red blood cell and platelet counts, haemoglobin concentration, mean corpuscular volume, mean corpuscular haemoglobin, and mean corpuscular haemoglobin concentration); differential leukocyte counts; blood biochemistry (plasma total protein, albumin, cholesterol, and glucose); mucosal histology (epithelial cell height, mucosal thickness, and villus height); and bacterial translocation to extra-gut tissues (blood, liver, spleen, kidney and mesenteric lymph nodes). DNA finger printing techniques were used to identify any viable bacterial strains recovered from these tissues. The results demonstrated that 4 weeks consumption of these LAB strains had no adverse effects on animals' general health status, haematology, blood biochemistry, gut mucosal histology parameters, or the incidence of bacterial translocation. A few viable LAB cells were recovered from the tissues of animals in both control and test groups, but DNA fingerprinting did not identify any of these as the inoculated strains. The results obtained in this study suggest that the potentially probiotic LAB strains HN001, HN017, and HN019 are non-toxic for mice and are therefore likely to be safe for human use.

Morphological changes in the oesophagus of newborn pigs: effects of age, diet and oral insulin-like growth factor I (IGF-I) or IGF-II.

It has been reported in the literature that the stomach and the intestine in newborns undergo profound growth and functional maturation during the immediate postnatal period and diet ingestion has a significant impact on these changes. The present paper examines oesophageal development in newborn pigs during the first three postnatal days and the effects of diet and oral insulin-like growth factor I (IGF-I) or IGF-II on oesophageal morphology. It was observed that marked changes, including reduction in thickness of the epithelium, accelerated proliferation and migration of basal epithelial cells and increased accumulation of mucus in the glandular cells, occurred during the first postnatal day following onset of natural suckling. Bottle-feeding with various liquid diets (i.e. porcine colostrum, bovine colostrum, bovine milk, and infant milk formula), induced marked morphological changes which were similar to those induced by natural suckling. However, bottle-feeding with water did not result in marked reduction in the thickness of the epithelium nor did it accelerate basal epithelial cell proliferation and migration. Oral IGF-I, but not IGF-II, increased basal epithelial cell proliferation up to 81%. Owing to a large inter-animal variation, the increment did not reach a significant level (P = 0.071). The results suggest that chemical constituents in the diet and physical stimulation of food ingestion, which cause sloughing off of luminal surface tissue, are two major stimuli or epithelial cell proliferation in the new born oesophagus.

Impact of intrauterine growth retardation on the gastrointestinal tract and the pancreas in newborn pigs.

Effects of intrauterine growth retardation (IUGR) on the gastrointestinal tract and the pancreas were examined using a model of piglets with naturally occurring IUGR. IUGR was associated with significant reductions in gastrointestinal and pancreatic tissue weights. The reduction in tissue weight in the gastrointestinal tract was due to lowered cell numbers and was proportionate to the body as a whole. In contrast to the gastrointestinal tract, the reduction in the pancreas was due to both lowered cell numbers and smaller cell size and was disproportionately greater than the body as a whole. IUGR was associated with precocious occurrence of maltase and sucrase activities in the small-intestinal mucosa. The exocrine cells of the pancreas in the animals with IUGR appeared immature when compared with those in normal piglets. Further studies are required to examine the postnatal effects of IUGR on the gastrointestinal tract, the pancreas, and somatic development.

Effects of oral IGF-I or IGF-II on digestive organ growth in newborn piglets.

To study whether colostrum-borne growth factors are responsible for the rapid GI tissue growth in naturally suckled newborn animals, newborn unsuckled piglets were bottle-fed for 24 h with infant milk formula with or without addition of 2 micrograms/ml of recombinant human insulin-like growth factor-I (IGF-I) or insulin-like growth factor-II (IGF-II), a level which approximated that of porcine colostrum. The animals were then sacrificed for measurements of their digestive organ weights and contents of protein, RNA and DNA in the organs. The treatment with IGF-I or IGF-II failed to show any significant effect on the weight of the esophagus, stomach, small intestine, large intestine, mandibular glands, kidneys and the spleen, and had no effects on the contents of protein, RNA and DNA in the small intestinal mucosa, the liver and the spleen. However, piglets fed with infant formula containing IGF-I (n = 7) or IGF-II (n = 7) had a heavier pancreas (p < 0.05) compared to formula-fed controls (n = 7). The DNA content in the stomach and the pancreas were greater in animals treated with IGF-I or IGF-II than in controls. Using a cell labelling technique it was shown that both IGF-I and IGF-II stimulated cell proliferation in the small intestinal crypts. The results indicate that the substantial GI tissue growth reported in newborn animals is unlikely due to colostrum-borne IGF-I or IGF-II alone. On the other hand the study does suggest that oral IGF-I and IGF-II are capable of stimulating cell proliferation in the GI tract.

Growth and morphological changes in the small and the large intestine in piglets during the first three days after birth.

Growth and morphological changes in the small and the large intestine of piglets were examined during the first three days after birth. There was a 72% increase in small intestinal weight, virtually all of which occurred during the first day and was due primarily to a 115% increase in the weight of the mucosa. Associated with the tissue weight gain there was a 24% increase in small intestinal length, a 15% increase in small intestinal diameter, a 33-90% increase in villus height and a 14-51% increase in villus diameter, during the first day. The cellular population in the small intestinal mucosa, as indicated by its DNA content, increased progressively with age, and at three days had increased by 84-154%. The percentage increase in mucosal DNA content was highest in the duodenum, intermediate in the jejunum and lowest in the ileum. Histological features and tissue protein contents revealed a transient epithelial cellular swelling related to intracellular accumulation of protein on the first day. Protein accumulation was evident in the jejunum and ileum but not in the duodenum. The positions of the nuclei in the epithelial cells suggested that on the first day protein absorption was at a more advanced stage in the jejunum and the proximal ileum than in the distal ileum. Large intestinal weight increased by 33% during the first day and had doubled by the third day, and this weight gain was due to both mucosal and non-mucosal tissue growth. Villus-like structures were observed in the caecum and the proximal colon in piglets at birth and one day after birth but not in piglets three days after birth. It is speculated that such villus-like structures may have a functional significance during the transition to complete dependence on oral nutrition in newborns.

Bovine ceroid-lipofuscinosis: pathology of blindness.

Five Devon cattle with suspected ceroid-lipofuscinosis and aged between 19 and 39 months of age were humanely slaughtered and subjected to post-mortem examination. There was severe atrophy of the cerebrum, particularly of the occipital cortex. Microscopy also showed severe atrophy of the retina with complete loss of photoreceptor cells, even in the youngest animal examined. Histopathologically the disease was characterised by accumulation of a fluorescent lipopigment in neurones, including those of the retina and a severe astrocytosis. The disease, which is characterised by the accumulation of subunit c of mitochondrial ATP synthase, is similar to that extensively described in South Hampshire sheep except that the retinal lesions were more severe. In contrast, tremors were not noted in the cattle. The clinical history and similarity to the disease in sheep and other species indicated inheritance was as an autosomal recessive trait.

Growth and morphological changes in the stomach of newborn pigs during the first three days after birth.

Growth and morphological changes in the stomach of newborn pigs were examined during the first 3 days after birth. The stomach grew disproportionately faster than the body as a whole during this period. The growth was due to hyperplasia and hypertrophy during the first day and mainly to hyperplasia thereafter as gastric DNA content increased progressively after birth, and the protein:DNA and RNA:DNA ratios increased only on the first day. Histological and morphometric analyses revealed that the growth was more pronounced in the gastric body region than in the cardiac and pyloric regions, and more pronounced in the mucosal layer than in other layers. The percentage of mucosal volume occupied by parietal cells (volume density) and the number of parietal cells per unit volume of gastric mucosa (numerical density) increased significantly 3 days after birth in the cardiac and body regions, but not in the pyloric region, of the stomach. The observed morphological changes coincide with the known pattern of functional maturation during the immediate postnatal period. It is suggested that a high level of circulating gastrin and oral ingestion of milk-derived growth factors in the newborn pig contribute to these changes.

Development of an immunocytochemical procedure to detect adenoviral antigens in chicken tissues.

An immunocytochemical technique utilizing an avidin-biotin peroxidase complex was developed to detect viral antigens in various tissues following oral administration of a locally isolated serotype 8 avian adenovirus (AAV) in specific pathogen-free (SPF) chickens. A strong color reaction was obtained with tissues from infected birds that contained a minimal amount of AAV antigens as determined by an indirect enzyme-linked immunosorbent assay. No reaction was detected in sections of tissues obtained from SPF chickens, and the reactivity with infected tissues could be removed by prior absorption of the primary antibody with purified AAV. A group-specific antigen common to the 12 serotypes of AAV was demonstrated by this technique. Because of the high sensitivity and broad-spectrum reactivity, this technique could be useful for studying the pathogenesis and laboratory diagnosis of inclusion body hepatitis caused by several serotypes of AAV.

An immunocytochemical method for studying patterns of cell proliferation in the wool follicle.

The identification of cell proliferation sites in the wool follicle bulbs of the skin of New Zealand Romney sheep was investigated with two immunocytochemical techniques. These methods were based on the in vivo labelling of DNA synthesising follicle matrix cells with the thymidine analogue 5-bromo-2'-deoxyuridine (BrdU) and a surgical preparation of the skin on the lateral abdominal flank of the sheep. Using a monoclonal antibody to BrdU, an indirect immunoenzyme method and a biotin-streptavidin method were compared for specificity and sensitivity in detecting replicating bulb matrix cells which had incorporated infused BrdU during the S-phase of the cell cycle. The immunocytochemical results for both methods showed a black-brown staining reaction of cell nuclei entering mitosis. The biotin-streptavidin method proved to be more highly specific and sensitive than the immunoenzyme technique. The immunocytochemical detection of cell cycle S-phase is highly suited for studying cell proliferation sites and cytokinetics in wool follicle bulbs and in other mitotically active tissues. Immunocytochemical detection of mitotically active cells has the advantages of high specificity, cost efficiency and rapidity and may be an alternative to methods employing metaphase arresting agents like colchicine or autoradiography.

Ceroid-lipofuscinosis (Batten's disease): pathogenesis and sequential neuropathological changes in the ovine model.

A sequential morphological study of ovine ceroid-lipofuscinosis showed that brains of affected lambs were normal at birth, grew until four months of age but then atrophied. Laminar necrosis of cerebral cortex was noted at 10 weeks, occurring first in the parietal area, then spreading to involve frontal and occipital areas while the temporal lobe was least and last affected. With progression of the disease, the laminar pattern was lost. Neuronal necrosis was accompanied by a severe astrocytosis. The granular and multilamellar storage cytosomes increased in size with age. Their structure was interpreted as paracrystalline in which repeating molecules of the dominantly stored lipid binding subunit of mitochondrial ATP synthase interact with neutral lipids and phospholipids. Abnormal cytosomes in neurons of lamb fetuses and a neonate were interpreted as early lesions which contained whorls or stacks of bilayered membrane as well as the more complex multilamellar material. The underlying anomaly leading to the storage of the lipid binding subunit of mitochondrial ATPsynthase remains to be defined. However, it is noted that this disease should be regarded as a lipid binding protein proteinosis or alternatively as a proteolipid proteinosis.

An estimate of the mean length of collagen fibrils in rat tail-tendon as a function of age.

A theoretical expression has been derived for the mean collagen fibril length in tendon based on the assumption that collagen fibrils originate in cell surface invaginations and terminate either at some remote cell surface or another collagen fibril bundle. The expression thus determined requires knowledge of the effective lengths of the fibrocytes (or fibrocyte assemblies) and the cellular content of the tendon. Both of these parameters have been measured experimentally as a function of age for rat-tail tendon using a combined light microscope and electron microscope approach. The results obtained for immature tendon suggest that the mean collagen fibril length is at least equal to the critical length required to maintain the appropriate tensile properties. In the most mature tissue studied, however, the mean-collagen fibril length is in excess of 100 times the critical length.

Age-related functional changes in the follicle-associated epithelium of the bursa of Fabricius in Shaver cockerels.

A study of the age-related functions of immunologically important components of the bursa of Fabricius in Shaver cockerels showed that endocytosis of carbon particles by the specialised follicle-associated epithelium was at a high level from hatching until 5 weeks of age and thereafter declined until at 18 weeks it could no longer be detected. The follicle-associated epithelium had marked non-specific esterase activity during the first 15 weeks of life as determined by a standard acid alpha-naphthyl acetate esterase method. The absolute weight of the bursa was at a maximum at 9 to 10 weeks. Involution began before 14 weeks and was complete by 22 weeks. The results indicate that the critical period for the bursa in regard to acquiring immunity from either local vaccination or environmental challenge is likely to be within the first five weeks of life.

Light microscopic observations of an intraepithelial granular cell type in the bovine parotid gland.

Bovine parotid gland intralobular ducts were frequently found to contain an intraepithelial granular cell type, adjacent to the basement membrane. Granular cells were non-migratory and polymorphic in appearance, with strong cytoplasmic basophilia and acidophil metachromatic granules. Such cells were not found in the bovine mandibular and sublingual glands. It is proposed that the granular cell may be similar or identical to the globule leucocyte cell type and functionally associated with the parotid gland. A possible role for parotid gland granular cells in ruminant mucosal immunity is also discussed.