The effect of dietary faba bean and non-starch polysaccharide degrading enzymes on the growth performance and gut physiology of young turkeys.

The aim of this study was to investigate the effect of dietary replacement of soya bean meal (SBM) with faba bean (FB) and a blend of non-starch polysaccharide (NSP) degrading enzymes on the gastrointestinal function, growth performance and welfare of young turkeys (1 to 56 days of age). An experiment with a 2×2 factorial design was performed to compare the efficacy of four diets: a SBM-based diet and a diet containing FB, with and without enzyme supplementation (C, FB, CE and FBE, respectively). In comparison with groups C, higher dry matter content and lower viscosity of the small intestinal digesta were noted in groups FB. The content of short-chain fatty acids (SCFAs) in the small intestinal digesta was higher in groups FB, but SCFA concentrations in the caecal digesta were comparable in groups C and FB. In comparison with control groups, similar BW gains, higher feed conversion ratio (FCR), higher dry matter content of excreta and milder symptoms of footpad dermatitis (FPD) were noted in groups FB. Enzyme supplementation increased the concentrations of acetate, butyrate and total SCFAs, but it did not increase the SCFA pool in the caecal digesta. The enzymatic preparation significantly improved FCR, reduced excreta hydration and the severity of FPD in turkeys. It can be concluded that in comparison with the SBM-based diet, the diet containing 30% of FB enables to achieve comparable BW gains accompanied by lower feed efficiency during the first 8 weeks of rearing. Non-starch polysaccharide-degrading enzymes can be used to improve the nutritional value of diets for young turkeys, but more desirable results of enzyme supplementation were noted in the SBM-based diet than in the FB-based diet.

Adrenergic control of pinealocyte chondriome - an in vitro study.

Norepinephrine released from sympathetic innervation plays the main role in the regulation of melatonin secretion in mammalian pinealocytes. The present study was conducted for the following reasons: 1) to establish whether the pinealocyte chondriome is controlled by norepinephrine, 2) to determine the effect of adrenergic stimulation on mitochondria, and 3) to characterize adrenoceptors involved in the regulation of the chondriome. The static organ culture of the pineal gland was used. The explants were incubated for 5 consecutive days in control medium and between 20:00 and 08:00 in medium with the presence of 10 µM norepinephrine - adrenergic agonist; isoproterenol - beta-adrenoceptor agonist; cirazoline, methoxamine, M-6364 - alfa1 - adrenoceptors agonists or PMA - activator of PKC. The explants were then subjected to ultrastructural examination and morphometric analysis. The incubation of explants in the presence of norepinephrine or isoproterenol caused a decrease in the relative volume and the numerical density of mitochondria and induced an increase in the percentage of free mitochondria in pinealocytes. Significant changes in these parameters were not observed after treatment with methoxamine, cirazoline, M-6463 and PMA. The results obtained show that the chondriome of pig pinealocytes is controlled by norepinephrine acting via beta-adrenoceptors. Adrenergic stimulation, repeated for five consecutive days of organ culture, causes a decrease in the number of mitochondria and a shift in the distribution of mitochondria from the form of networks and filaments into the form of single particles. This indicates the intensive remodeling of the mitochondria network, which is closely linked to the metabolic status of the cell.

Regulation of melatonin secretion in the pineal organ of the domestic duck--an in vitro study.

The aim of study was to determine the mechanisms regulating melatonin secretion in the pineal organs of 1-day-old and 9-month-old domestic ducks. The pineals were cultured in a superfusion system under different light conditions. Additionally, some explants were treated with norepinephrine. The pineal glands of 1-day-old ducks released melatonin in a well-entrained, regular rhythm during incubation under a 12 hrs light:12 hrs dark cycle and adjusted their secretory activity to a reversed 12 hrs dark:12 hrs light cycle within 2 days. In contrast, the diurnal changes in melatonin secretion from the pineals of 9-month-old ducks were largely irregular and the adaptation to a reversed cycle lasted 3 days. The pineal organs of nestling and adult ducks incubated in a continuous light or darkness secreted melatonin in a circadian rhythm. The treatment with norepinephrine during photophases of a light-dark cycle resulted in: 1) a precise adjustment of melatonin secretion rhythm to the presence of this catecholamine in the culture medium, 2) a very high amplitude of the rhythm, 3) a rapid adaptation of the pineal secretory activity to a reversed light-dark cycle. The effects of norepinephrine were similar in the pineal organs of nestlings and adults. In conclusion, melatonin secretion in the duck pineal organ is controlled by three main mechanisms: the direct photoreception, the endogenous generator and the noradrenergic transmission. The efficiency of intra-pineal, photosensitivity-based regulatory mechanism is markedly lower in adult than in nestling individuals.

Neuropeptide Y as a presynaptic modulator of norepinephrine release from the sympathetic nerve fibers in the pig pineal gland.

Norepinephrine (NE) released from the sympathetic nerve endings is the main neurotransmitter controlling melatonin synthesis in the mammalian pineal gland. Although neuropeptide Y (NPY) co-exists with NE in the pineal sympathetic nerve fibers it also occurs in a population of non-adrenergic nerve fibers located in this gland. The role of NPY in pineal physiology is still enigmatic. The present study characterizes the effect of NPY on the depolarization-evoked 3H-NE release from the pig pineal explants. The explants of the pig pineal gland were loaded with 3H-NE in the presence of pargyline and superfused with Tyrode medium. They were exposed twice to the modified Tyrode medium containing 60 mM of K+ to evoke the 3H-NE release via depolarization. NPY, specific agonists of Y1- and Y2- receptors and pharmacologically active ligands of α2-adrenoceptors were added to the medium before and during the second depolarization. The radioactivity was measured in medium fractions collected every 2 minutes during the superfusion. NPY (0.1-10 µM) significantly decreased the depolarization-induced 3H-NE release. Similar effect was observed after the treatment with Y2-agonist: NPY13-36, but not with Y1-agonist: [Leu31,Pro34]-NPY. The tritium overflow was lower in the explants exposed to the 5 µM NPY and 1 µM rauwolscine than to rauwolscine only. The effects of 5 µM NPY and 0.05 µM UK 14,304 on the depolarization-evoked 3H-NE release were additive. The results show that NPY is involved in the regulation of NE release from the sympathetic terminals in the pig pineal gland, inhibiting this process via Y2-receptors.

Gastrointestinal morphology and function in turkeys fed diets diluted with whole grain wheat.

The aim of this study was to verify the hypothesis that moderate dilution of turkey diets with whole grain wheat (up to 22.5%) improves gastrointestinal function, thus contributing to sustained growth performance. A total of 900 male turkeys were allocated to 5 dietary treatments: basal diet (BD), BD diluted with low levels of ground and pelleted wheat or whole wheat, and BD diluted with high levels of ground and pelleted wheat or whole wheat (GH and WH, respectively). At successive stages of the experiment, diets containing 5 and 15%, 7.5 and 17.5%, 10 and 20%, 12.5 and 22.5% of wheat in different physical form were fed to turkeys aged 5 to 8, 9 to 12, 13 to 16, and 17 to 18 wk, divided into subgroups with low and high dietary levels of wheat, respectively. Dietary dilution with different levels of wheat did not reduce the BW gain of turkeys and did not deteriorate feed conversion despite a decrease in the amino acid content of diets. In comparison with BD, diet GH increased the thickness of the muscularis externa and the depth of cecal crypts (P < 0.05), increased Bacteria domain, and decreased Salmonella counts (P < 0.001), increased butyric acid concentrations, and decreased the levels of putrefactive compounds in the cecal digesta (P < 0.001). In comparison with BD, diet WH improved feed efficiency (P = 0.002) and contributed to significant (P < 0.01) changes in the parameters of gastrointestinal function, including lower pH of gizzard digesta, increased depth of jejunal crypts, lower ammonia concentrations, and lower pH of intestinal digesta, increased Bacteria and Bifidobacterium counts, decreased Salmonella counts, and higher concentrations of butyric acid and total short-chain fatty acids. In comparison with diet GH, diet WH enhanced the activity of bacterial α-glucosidase and ß-glucosidase as well as α-galactosidase, reduced the activity of ß-glucuronidase, and increased total fatty acid concentrations, mainly acetic acid levels, in the cecal digesta (P < 0.01). The observed changes indicate that diet supplementation with wheat grain enhances gastrointestinal function, thus improving feed conversion in turkeys.

The low doses effect of experimental zearalenone (ZEN) intoxication on the presence of Ca2+ in selected ovarian cells from pre-pubertal bitches.

The objective of this study was to determine the effect of 42-day ZEN intoxication on the presence of Ca2+ in selected ovarian cells from beagle bitches, using the potassium pyroantimonate (PPA) method. Samples were collected from 30 clinically healthy, pre-pubertal, genetically homogeneous animals. The bitches were divided into three groups of 10 animals each: experimental group I--50 microg ZEN/kg BW (100% NOAEL) administered once daily per os; experimental group II--75 microg ZEN/kg BW (150% NOAEL) administered once dailyper os; control group--placebo containing no ZEN administered per os. An electron microscopic analysis revealed that cells died due to apoptosis, depending on the ZEN dose and the type of cells exposed to intoxication. Lower ZEN doses led to apoptosis-like changes in the cells. Cell death was a consequence of excess Ca2+ accumulation in the mitochondria, followed by cell dysfunction and a decrease in or the absence of mitochondrial metabolic activity in oocytes, follicle cells and interstitial cells in experimental bitches.

The influence of methisoprinol administered in ovo on the morphological structure of the spleen in turkeys.

This study analyzed the effect of a synthetic, low-toxic immunomodulator - methisoprinol - administered in ovo on the morphological structure of the spleen in turkeys. Experiments were conducted on three groups of 5-day-old BUT 9 turkeys (35 birds in each group) hatched from eggs which, on day 26 of incubation, had been administered methisoprinol (VetAgro, Lublin, Poland) in ovo in a dose of 5 mg (group I) or 20 mg per egg (group II). Poults hatched from eggs administered a physiological solution of NaCl in a dose of 0.1 ml per egg in ovo served as a control (group III). Samples of the spleen were collected from 5 birds selected at random from a group of decapitated 5-day-old poults and the prepared 7 fm-paraffin sections were stained with HE. A morphometric analysis of the germinal centres of the white pulp of the spleen was conducted by subjecting pictures taken with an optical microscope to a Digital Image Analysis using Axio Vision software (by Zeiss). The study demonstrated that in terms of the morphological structure, the spleen of the poults hatched from eggs administered 5 mg of methisoprinol (group I) did not differ considerably from the spleen of the control birds. In turn, spleens of the poults hatched from eggs administered 20 mg of methisoprinol per embryo were characterized by distinctively developed red pulp and within the area of the white pulp by distinct cortical section containing numerous lymphocytes. In spleens of the poults from this group, the morphometric examination also demonstrated a higher number of germinal centres of the white pulp as compared to their number in spleens of the birds from the other groups.

Calcium concrements in the pineal gland of the Arctic fox (Vulpes lagopus) and their relationship to pinealocytes, glial cells and type I and III collagen fibers.

The aim of the present study was to analyze the presence and morphology of the pineal concretions in the Arctic fox and their relationship to pinealocytes, glial cells and collagen fibers. Pineals collected from 7-8 month-old and 3-4 year-old foxes (6 in each age-group) were investigated. Sections of the glands were stained with HE, Mallory's method and alizarin red S as well as subjected to a combined procedure involving immunofluorescent staining with antibodies against antigen S, glial fibril acid protein (GFAP), type I and III collagen and histochemical staining with alizarin red S. The pineal concretions were found in 2 of 6 investigated Arctic foxes aged 3 years and they were not observed in animals aged 7-8 months. The acervuli were present in the parenchyma and the connective tissue septa. They were more numerous in the distal part than in the proximal part of the gland. The acervuli stained with alizarin red S revealed an intensive red fluorescence, what enabled the use of this compound in a combined histochemical-immunofluorescent procedure. A majority of cells in the fox pineal showed positive staining with antibodies against antigen S, a marker of pinealocytes. GFAP-positive cells were especially numerous in the proximal part of the gland. Both antigen S- and GFAP-positive cells were frequently observed close to the concrements. Collagen fibers of type I and III were found in the capsule, connective tissue septa and vessels. Immunoreactive fibers did not form any capsules or basket-like structures surrounding the concrements.

Pineal concretions in turkey (Meleagris gallopavo) as a result of collagen-mediated calcification.

The intra-pineal calcification is a well-known phenomenon in mammals, however it is almost completely unknown in birds. The aim of the present work was to analyze morphology and genesis of the pineal concretions in the turkey. The studies were performed on the pineals collected from one-year-old turkeys (Meleagris gallopavo). In addition to standard morphological methods, the alizarin red S and potassium pyroantimonate methods were employed for localization of calcium at the light and electron microscopy level. In light microscopy, calcified concretions with diameters from 300 microm to 2 mm and quantities from 3 to 6 per gland were observed in all the examined pineals. They were stained red with alizarin S and showed the presence of collagen in Mallory's staining. Two types of cells were noted inside the concretion: polygonal and elongated ones. Using electron microscopy, three parts were distinguished within the calcification area. The peripheral part contained densely packed collagen fibrils, some elongated cells and numerous pyroantimonate precipitates demonstrating the presence of calcium ions. In the intermediate part, the fibrils were covered by almost continuous sheets of pyroantimonate precipitates and fused side by side. The central part showed an appearance of calcified hard tissue and contained some polygonal (osteocyte-like) cells. The obtained data demonstrated that the formation of the pineal concretions in the turkey is associated with the mineralization of collagen. This process is completely different from the mechanisms responsible for the formation of the concretions in the mammalian pineal.

Effect of diets with different contents of soybean alpha-galactosides and crude fibre on modification of duodenal microstructure and selected parameters of nutrient utilization in young turkeys.

The aim of the present study was to assess the physiological response of growing turkeys' duodenal surface to dietary replacement of a common dietary component--soybean meal (SBM) with a soy protein isolate (SPI), as this treatment was associated with almost complete removal of alpha-galactosides from the diet (from 2.44 to 0.15%). Additionally, the utilization of selected dietary ingredients upon dietary treatments was recorded. Effects of raffinose-family oligosaccharides were assessed as well in low- and high-fibre dietary environment (3.5 and 5.3% of crude fibre, respectively). This study revealed that the duodenal morphological parameters were differently affected by dietary treatments at different production stages, i.e. at 4 and 8 weeks of life. Although villus height/crypt depth ratio (VCR) was insignificantly decreased by high-alpha-galactoside treatment in younger 4-week birds, the presence of these oligosaccharides in the diet positively influenced the VCR index in 8-week turkeys. A similar tendency was observed when calcium retention was considered. Different contents of dietary crude fibre affected the physiological action of alpha-galactosides, including duodenal crypts depth and phosphorus retention in the 4-week birds, as well as duodenal goblet cells number and nitrogen utilization in the older turkeys. A high content of alpha-galactosides in the diet resulted in increased hydration of intestinal contents, but without a significant decline in the dry matter digestibility and utilization of nitrogen, calcium and phosphorus. Having in mind the development and physiology of the GIT, it may preliminary be concluded that in later production stages, total withdrawal of soybean alpha-galactosides from turkeys' diets does not seem to be nutritionally advisable.

Abnormal morphology of skeletal muscles in meat-type chickens--ultrastructural observations.

In order to determine the effect of different production systems on muscle ultrastructure in meat-type chickens, we examined m. gastrocnemius and m. pectoralis superficialis in two lines of chickens (Anak Titan and Isa 215) raised in three different technological systems (indoors in a conventional facility, indoors with limited outdoor access and outdoors with an umbrella roof). Our previous study showed some abnormalities in the histological structure of these muscles. We hypothesized that electron microscopy, having a strategic position in muscle examination, would provide insight into changes in muscle tissue revealed by light microscopy. The results of the study indicate that the muscles examined undergo ultrastructural alterations regardless of the muscle type, chicken line and production system. The abnormalities observed in the present study were found to affect many aspects of fiber ultrastructure impairing function of the myofibrillar apparatus--the structure of mitochondria and sarcoplasmic reticulum, defects of the sarcolemma as well as the appearance of remnants resulting from fiber disintegration. Abnormal responses were found primarily in myofibrils and mitochondria, and--to a lesser extent--in other structures of muscle fibers. We suggest that the majority of the changes observed may lead to muscle damage followed by pathology. The severity of these changes was particularly evident in the muscles of chickens of the Isa 215 line (highly selected) kept outdoors with an umbrella roof. The observations point to a dependence of the above changes on the line of chickens and rearing conditions. Therefore, the limited potential of highly selected broilers to adapt to different environmental conditions should be taken into account while selecting a new production technology.

Ultrastructural changes of ovarian follicle and corpus luteum after experimental zearalenone mycotoxicosis in bitch.

The aim of the study was to determine the influence of experimental zearalenone (ZEA) mycotoxicosis on the ultrastructure of ovaries in bitches receving zearalenone for 100 days per os in the anestrus phase. Experiment has been conducted on 9 sexually mature clinically healthy bitches. The sexually mature animals aged 1-3 years were in anestrus. The dogs have been divided into three groups: EI and EII, receiving ZEAper os in two dosages (25 microg/kg and 50 microg/kg b.w.) and control animals which received placebo per os. On the last day of experiment, ovariohysterectomy was performed in all the bitches and the uterine samples were submitted to ultrastructural analyses. The study has revealed that long lasting administration of higher dose of zearalenone causes ultrastructural changes in the granular layer of ovarian follicles, which showed due to their biological activity decrease. The changes in the granular layer, atrophy of intercellular connections and the appearance of intercellular spaces can stand for reduced activity of this layer. Changes in morphology of mitochondria, rough endoplasmic reticulum and Golgi apparatus sugest ZEA influence on cell metabolism and secretory processes. It seems therefore that in bitches and other monogastric animals, intoxications induced by this mycotoxin causes ultrastructural changes in granular cells of ovarian follicles.

Histology and ultrastructure of the pineal organ in the domestic goose.

The pineal organs of 14-week-old domestic geese were investigated with light and electron microscopy. The pineals consisted of a wide distal part and a narrow middle-proximal one. The glands were attached to the intercommissural region via the choroid plexus. The pineal parenchyma was formed by round or elongated follicles. The follicular wall was composed predominantly by cells immunoreactive with antibodies against hydroxyindolo-O-methyltransferase (HIOMT) or glial fibrillary acid protein (GFAP). They formed two or more layers. HIOMT-positive elements were represented by elongated cells bordering the follicular lumen and oval cells located in the external layer of the follicular wall. These cells were identified in ultrastructural studies as rudimentary-receptor pinealocytes and secretory pinealocytes, respectively. Among rudimentary-receptor pinealocytes two types of cells, designed as A and B, were distinguished due to structural differences. Type A cells extended through the whole follicular wall and showed regular stratified distribution of organelles in well-recognizable zones with rough endoplasmic reticulum, the Golgi apparatus and mitochondria. Type B cells, like type A pinealocytes, contacted the pineal lumen and showed polarity of their internal structure. However, they were markedly shorter than the cells of type A and lacked stratified distribution of organelles. Secretory pinealocytes contained irregularly dispersed organelles. A prominent feature of all types of goose pinealocytes was the presence of numerous dense core vesicles. The population of GFAP-positive cells consisted of ependymal-like supporting cells and astrocyte-like cells.

Postnatal development of the pineal gland in the goat (Capra hircus)--light and electron microscopy studies.

This study was conducted to investigate structural transformations of the goat pineal gland during postnatal development. The pineals of newborn, 4-week-, 4-month-, 1-year- and 3-year-old male goats were prepared for qualitative and quantitative investigations at both light and transmission electron microscopy levels. In the first 4 months after birth, the pinealocytes developed very intensively, which was connected with enlargement of their volume as well as distinct qualitative and quantitative transformations of their cytoplasmic organelles. Parallel to these changes, the pineal parenchyma underwent deep reorganization and the mosaic pattern, characteristic for newborns, disappeared. At the same time the meshwork of blood vessels supporting the goat pineal gland developed conspicuously. From fourth months to one year of postnatal life continuation of the growth of some pinealocyte organelles and very intensive development of all components of the pineal connective tissue were observed. In the three-year-old goats some decrease in the relative volume of most cytoplasmic structures of pinealocytes as well as increase in collagen fibers in stroma were noticed. The investigations established intensive growth of the goat pineal to the age of 4 months, which included mainly some increase in the pinealocyte and their structures. The pineal gland of 4-month- and 1-year-old goats showed morphological features which are considered as symptoms of high secretory activity. The structure of the pineal gland in 3-year-old animals and the changes observed both in their pinealocytes and connective tissue in comparison with 1-year-old individuals point to some decrease in the goat pineal activity in this age.

Distribution of NADPH-diaphorase activity in the pineal gland of the Turkey.

NADPH-diaphorase activity was histochemically demonstrated in the nerve fibers, neuronal-like cell bodies and in the endothelial cells of the vasculature in the pineal gland of the turkey. The nerve fibers were localized in the choroid plexus, connecting the pineal gland with the diencephalon as well as inside the pineal gland, where they formed basket-like structures around the pineal follicles. A group of neuronal-like cell bodies was observed in the proximal part of the gland. The positive staining was not observed in the pinealocytes of rudimentary-photoreceptor type and in the supporting cells.

Leu-enkephalin immunoreactivity in the pig pineal gland.

Leu-enkephalin-positive structures in the pig pineal gland were demonstrated immunohistochemically using mouse monoclonal antibody. The pineal glands were obtained from the newborn, 21-day and 7-month old female pig. The immunopositive nerve fibers were observed in the pineal gland as well as in the epithalamic areas. The leu-enkephalin-immunoreactive nerve fibers (single or forming small bundles) were localized mainly in the proximal part of the pineal and they were scarce in other parts. The localization of the fibers points to a central source of this innervation. The study did not show any age-dependent differences in the distribution and density of leu-ekephalin-positive nerve fibers.

Pituitary adenylate cyclase-activating polypeptide-immunoreactive (PACAP-IR) nerve fibers in the pig pineal gland.

The present study demonstrates the occurrence of PACAP-immunoreactive (PACAP-IR) nerve fibers in different compartments of the pig pineal gland, including glandular capsule (where they form a very dense network) and subependymal tissue close to the pineal recess (moderate to dense meshwork of varicose fibers). Furthermore, several varicose fibers penetrate from the capsule into the connective tissue septa and then into the parenchyma, where they form unequally distributed, fine network and, in some cases, basket-like structures around pinealocytes. Some of the PACAP-IR nerve fibers, observed both in the habenular and posterior epithalamic areas, extend to the pineal gland. PACAP-IR cells could be demonstrated neither in the pineal gland, nor in epithalamic areas.

Localization of NADPH-diaphorase activity in the pineal gland of the domestic pig.

The study showed the presence of NADPH-diaphorase containing structures in the pineal gland of the domestic pig. NADPH-diaphorase activity was found in the nerve fibers and in the endothelial cells of the vasculature. The nerve fibers were localized in the capsule, around the blood vessels as well as in the parenchyma. The positive staining was not observed in the pig pinealocytes.

Distribution of growth associated protein (GAP-43) immunoreactivity in nerve fibers supplying the pig pineal gland.

An immunohistochemical study of the pig pineal gland was carried out using monoclonal mouse antiserum against growth-associated protein GAP-43. The pineal glands were obtained from the 3, 5, 8 weeks old piglets. The immunopositive nerve fibers were observed in the pineal gland as well as in the habenular and the posterior comissural areas. They formed a dense network in the habenular area and the proximal part of the pineal gland. In the comissural area and in the apical part of the gland. single positive fibers were observed. The obtained results may suggest a difference in the plasticity of innervation between the particular regions of the pineal gland.

Immunohistochemical localization of substance P in the pineal gland of the domestic pig.

An immunohistochemical study of the pig pineal gland was carried out using polyclonal rabbit antiserum raised against substance P (SP). The pineal glands were taken from the newborn, 21-day- and 7-month-old female pigs. Immunoreactive nerve fibers were observed in the pineal gland as well as in the posterior commissure and habenular areas. The bundles of SP-immunoreactive fibers were also seen in the subependymal layer of the pineal tissue. The single SP-immunoreactive nerve fibers and few small bundles of nerve fibers were located with equal density throughout the pineal gland, in the connective tissue septa and in the parenchyma. SP-immunoreactive cell bodies were observed in the medial habenular nucleus. The obtained results point to this nucleus as one of the central sources of SP innervation in the pig pineal gland. The study did not show any differences in the distribution and the density of SP-immunoreactive nerve fibers between newborn, 21-day- and 7 month-old pigs.