Bioconservation of iron and enhancement of antioxidant and antibacterial properties of chicken gizzard protein hydrolysate fermented by Pediococcus acidilactici ATTC 8042.

BACKGROUND: The poultry industry is one of the fastest growing sectors, and it generates considerable quantities of chicken gizzards (CG) every day. However, due to their hard texture and high microbial load, and due to cultural beliefs, they are not preferred by consumers. Chicken gizzards are a substantial source of proteins, iron, and other nutrients, which can be used effectively to produce nutraceuticals, rich in peptides (antioxidants and antibacterial), bio-iron, essential free amino acids, and fatty acids vital for human health. RESULTS: Lactic acid fermentation of CG by Pediococcus acidilactici ATTC 8042 increased the antioxidant activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH), azino-bis (3-ethylbenzothiaziline-6-sulphonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) by up to 26 times compared with unfermented CG (P < 0.05). The amount of hydrolysis and solvents (ethanol and water) used for extracting protein hydrolysates significantly affected the antioxidant properties. Moreover, fermented CG showed a negligible reduction in bio-iron (2-3%) compared with heat-processed CG (85 °C for 15 min), in which bio-iron was reduced by up to 20.3% (P < 0.05). The presence of unsaturated fatty acids such as C20:4 and C22:4 n-6 indicated a low level of lipid oxidation. CONCLUSION: Fermented CG, with its reasonably high antioxidant and antibacterial activity, together with a substantial amount of bio-iron and other nutritional components can serve as a functional food or feed additive to reduce oxidative stress and to treat iron deficiency. © 2020 Society of Chemical Industry.

Food intake-related genes in chicken determined through combinatorial genome-wide association study and transcriptome analysis.

The chicken gizzard is the primary digestive and absorptive organ regulating food intake and metabolism. Body weight is a typical complex trait regulated by an interactive polygene network which is under the control of an interacting network of polygenes. To simplify these genotype-phenotype associations, the gizzard is a suitable target organ to preliminarily explore the mechanism underlying the regulation of chicken growth through controlled food intake. This study aimed to identify key food intake-related genes through combinatorial GWAS and transcriptome analysis. We performed GWAS of body weight in an F2 intercrossed population and transcriptional profiling analysis of gizzards from chickens with different body weight. We identified a major 10 Mb quantitative trait locus (QTL) on chromosome 1 and numerous minor QTL distributed among 24 chromosomes. Combining data regarding QTL and gizzard gene expression, two hub genes, MLNR and HTR2A, and a list of core genes with small effect were found to be associated with food intake. Furthermore, the neuroactive ligand-receptor interaction pathway was found to play a key role in regulating the appetite of chickens. The present results show the major-minor gene interactions in metabolic pathways and provide insights into the genetic architecture and gene regulation during food intake in chickens.

In vitro versus in situ evaluation of xylan hydrolysis into xylo-oligosaccharides in broiler chicken gastrointestinal tract.

Xylan hydrolysis into xylo-oligosaccharides (XOS) was evaluated both in the gizzard and ileum of broiler chickens, and by a 2-step in vitro digestion assay that simulated the pH, temperature and time period of the gastric and small intestine (SI) phases. Twelve dietary treatments with varying soluble and insoluble xylan levels, either with or without supplemental xylanase, were fed to broiler chickens (n = 576) for the in situ analysis, and were exposed to the in vitro assay. Relatedness of the two methods was strong for determination of XOS production in all dietary treatments for X5, X4, X3, X2 and X1, respectively, in both the gastric (r = 0.980, 0.853, 0.894, 0.870 and 0.951) and small intestine phase (r = 0.957, 0.923, 0.940, 0.970, 0.969) (P < 0.05). Consequently, the in vitro assay was used to illustrated the diversity of XOS production across different batches of wheat and barley in the presence of xylanase.

Elemental imbalance elicited by arsenic and copper exposures leads to oxidative stress and immunotoxicity in chicken gizzard, activating the protective effects of heat shock proteins.

Arsenic (As) and copper (Cu) are ubiquitous pollutants that pose a threat to the environment. Our aim is to study the underlying mechanisms by which As and Cu act on the chicken gizzard. In order to detect ionic disorders in chicken gizzard under chronic treatment with As3+ and/or Cu2+ and whether they can induce oxidative damage as well as immune disorders, 30 mg/kg arsenic trioxide (As2O3) and/or 300 mg/kg copper sulfate (CuSO4) were added to the chicken's basal diet. After 12 weeks of exposure, trace elements were found to have significant interference, accompanied by damage to the antioxidant system. In addition, As3+ and/or Cu2+ activated the nuclear factor kappa B (NF-κB), inducing severe inflammation. At the same time, damaged structural integrity which might be caused by inflammation was discovered after hematoxylin and eosin (H&E) staining. Moreover, symbolic Th1/Th2 (Th, helper T cell) drift was also observed in treatment groups, meaning that immune function is left to be affected, and the increment in heat shock proteins may be a self-protective mechanism of gizzard. Interestingly, we found that the damage to the gizzard of chicken was aggravated in a time-dependent manner, and the combined exposure was more pathogenic than the single exposure, of which the mechanism needs further exploration. Together, this work helps move us toward a better understanding of the molecular mechanisms that mediate the interactions between Cu excess and As3+ exposures and possible health consequences in susceptible species.

The disturbance of autophagy and apoptosis in the gizzard caused by copper and/or arsenic are related to mitochondrial kinetics.

As toxic elements when excessive, arsenic (As) and copper (Cu) are two naturally occurring elements that may be ingested by the organism at the same time. However, the precise damaged mechanism and the pathways that are activated by As and/or Cu is rarely researched in gizzard, a unique organ of birds. In this study, ultrastructural observations, TdT-mediated dUTP Nick-End Labeling, real-time quantitative PCR and Western blotting were performed to evaluate the toxic effects of chronic exposure to Cu2+ and/or arsenite on chicken gizzard. The results revealed that increased apoptosis and autophagy levels induced by Cu2+ and arsenite appeared to be independent of oxidative stress, which didn't have significant changes in different treatment groups at the same time point. Nevertheless, the redox balance gradually deviated with the extension of time. And increased mitochondrial division and decreased fusion were also caused by Cu2+ and arsenite. In conclusion, apoptosis and autophagy in gizzard induced by Cu2+ and/or arsenite, at least, strongly linked with the disruption of mitochondrial homeostasis. Our study showed that the combination of Cu2+ and arsenite produces stronger toxicity. The results of this study can serve as a reference for agicultural feeding and environmental protection, that is, to avoid the combined exposure of Cu2+ and arsenite to prevent greater economic losses and health risks.

Selenium Deficiency Induced Injury in Chicken Muscular Stomach by Downregulating Selenoproteins.

The aim of the present study was to examine the effect of selenium (Se) deficiency on the expression of selenoproteins in chicken muscular stomach and to detect the correlation of selenoproteins with muscular stomach injuries. One-day-old broiler chickens were maintained for 55 days on a normal diet (0.2 mg/kg) or a Se-deficient diet (0.033 mg Se/kg). The expression levels of 25 selenoproteins, heat shock proteins (HSPs), and inflammatory factors were then examined by real-time PCR. Following this, the correlation between selenoproteins, HSPs, and inflammatory factors was analyzed by principal component analysis (PCA). The results showed that Se deficiency decreased the expression of 25 selenoproteins (P < 0.05), but increased the expression of HSP27, HSP40, HSP60, HSP70, and HSP90, and NF-κB, iNOS, TNF-α, COX-2, and HO-1 (P < 0.05). Selenoproteins showed a high negative correlation with HSPs and inflammatory factors. Thus, the results suggested that Se deficiency induced muscular stomach injuries by decreasing the expression of selenoproteins. In addition, selenoproteins play an important role in regulating HSPs and inflammatory response. The muscular stomach is a key target of Se deficiency and may play a special role in response to Se deficiency.

Identification of the gene product recognized by monoclonal antibody GIIF3.

The chicken as a research model has a disadvantage compared with the mouse and the human because of the low number of available antibodies against gene products of interest. The goal of this study was to identify the antigen recognized by monoclonal antibody (mAb) GIIF3, which is a 42 kDa protein that appears in follicle-associated epithelium of the guinea hen as well as in different muscle types during chicken embryonic development. The 42 kDa protein, immunoprecipitated from chicken gizzard protein lysates, was evaluated by mass spectrometry. Mass spectrometry analysis revealed peptides specific for the chicken ß- or γ-actin isoforms. The mAb GIIF3 can be used as a new research tool for smooth muscle cell and bursa of Fabricius developmental studies.

Influence of superdoses of a novel microbial phytase on growth performance, tibia ash, and gizzard phytate and inositol in young broilers.

An experiment was conducted to evaluate the influence of a novel microbial phytase on performance, tibia ash, and the content of phytate, phytate esters, and inositol in the gizzard of young broilers. Male Cobb 500 broilers (n = 1,680) were fed 1 of 7 experimental diets: positive control (PC) formulated to meet or exceed nutrient recommendations; PC plus dicalcium phosphate (PC+DCP) formulated to provide Ca and P at 0.10% above the PC; PC plus 500 U/kg of microbial phytase (PC+500); negative control (NC) with Ca and P reduced from the PC by 0.16% and 0.15%, respectively; and the NC plus phytase at 500 (NC+500), 1,000 (NC+1,000), or 1,500 (NC+1,500) U/kg. Diets were fed in crumbled form to 20 birds/pen and 12 replicate pens/diet from d 0 to 21. On d 21, 4 birds/pen were euthanized for collection of right tibias and gizzard digesta for determination of tibia ash and gizzard phytate. In general, broilers fed the NC diet had reduced (P ≤ 0.05) feed intake and BW gain compared with broilers fed diets supplemented with phytase, but not different than the PC or PC+DCP. Phytase supplementation in the NC diet improved (P ≤ 0.05) BW gain comparable with or above that of the PC. Feed conversion ratio was improved in broilers fed the NC+1,000 or NC+1,500 compared with broilers fed all other diets. Tibia ash was reduced (P ≤ 0.05) in broilers fed the NC compared with broilers fed all other diets, and phytase supplementation improved tibia ash comparable with the PC. Phytase supplementation reduced (P ≤ 0.05) phytate (inositol hexa-phosphate) concentration in the gizzard. Inositol concentration in the gizzard was higher (P ≤ 0.05) in birds fed NC+1,000 or NC+1,500 compared with all other diets and this was correlated with growth performance (P ≤ 0.05) rather than tibia ash (P > 0.05). Improvements in feed conversion ratio associated with superdoses of phytase may be attributed to phytate destruction and the provision of inositol.

Comparison of atrogin-1/MAFbx mRNA expression in the gizzards of egg- and meat-type chickens.

We examined atrogin-1/MAFbx mRNA expression in the smooth muscle of gizzards from egg- and meat-type chickens. Gizzard weight relative to body weight was significantly lower in the meat-type chickens than in the egg-type at 14 d of age. In contrast, the level of atrogin-1/MAFbx mRNA in the gizzard was significantly higher in the meat-type chickens than in the egg-type chickens. Thus atrogin-1/MAFbx mRNA expression in the smooth muscle of the gizzard was higher in meat-type chickens than in egg-type chickens, in contrast to its expression in the skeletal muscles.

Lead concentrations in sediments and blue-winged teals (Anas discors) from El Palmar State Reserve, Yucatan, Mexico.

Reserve regulations at El Palmar State Reserve, Yucatan, Mexico, prohibit the use of lead (Pb) shot, but hunters continue to use it, and no enforcement is implemented. Pb was quantified in sediments and in blue-winged teal Anas discors. No shot pellets were found in the sediment samples, nor were differences in sediment Pb concentrations observed within the reserve between popular hunting sites and those no longer used for hunting. However, there were differences between the hunting sites and sediments from an adjacent area where hunting is prohibited. Average Pb concentrations were highest at hunting entrances (15.69 ± 18.69 mg/kg) and lowest at decoy locations (5.24 ± 4.84 mg/kg). These averages are lower than the lowest effects level (31 mg/kg), although 10 samples exceeded this level. Pb-shot prevalence in gizzards was 4.88% (n = 41). Pb levels exceeded 5.0 mg/kg dry weight in one or more of the tested tissues (liver, gizzard, and bone) in 14 (34.14%; 7 female, 7 male; 11 adult, 3 juvenile) of the total birds. Bird weight, sex, and age had no effect on Pb concentration. Hunting using Pb shot in the reserve clearly affects Pb levels in sediments and in A. discors that winter there.

Expression pattern of the homeotic gene Bapx1 during early chick gastrointestinal tract development.

Regulation of the Bone Morphogenetic Protein (BMP) signaling pathway is essential for the normal development of vertebrate gastrointestinal (GI) tract, but also for the differentiation of the digestive mesenchymal layer into smooth muscles and submucosal layer. Different studies demonstrated that Bapx1 (for bagpipe homeobox homolog 1) negatively regulates the BMP pathway, but its precise expression pattern during the development and the differentiation of the GI tract mesenchyme actually remains to be examined. Here, we present the spatio-temporal expression profile of Bapx1 in the chick GI tract. We show that Bapx1 is first expressed in the undifferentiated mesenchyme of the gizzard and the colon. After the differentiation of the digestive mesenchyme, we found Bapx1 strongly expressed in the gizzard smooth muscle and in the submucosa layer of the colon. This expression pattern provides new insights into the roles of Bapx1 during the regionalization of the GI tract and the differentiation of the digestive mesenchyme of the colon and the stomach.

µ-Calpain is involved in the postmortem proteolysis of gizzard smooth muscle.

Postmortem changes in proteins that have been implicated in affecting muscle integrity were examined in goose (GG) and duck (DG) gizzard smooth muscle stored at 5°C. GG and DG smooth muscles were sampled at 0, 1, 3 and 7 day of storage. The pH was approximately 7 in both GG and DG samples during postmortem storage. Casein zymograms showed that 0-day µ-calpain activity was higher (p<0.05) in GG than in DG samples. As postmortem time progressed, µ-calpain was activated and autolyzed more extensively in GG than in DG samples. However, µ/m-calpain remained relatively stable in both samples. Western blots indicated that postmortem desmin degradation was more rapid in GG than in DG samples. In contrast, α-actinin remained nearly unchanged in both samples. Therefore, our results suggest that µ-calpain has an important role in the postmortem proteolysis of gizzard smooth muscle.

Molluscan smooth catch muscle contains calponin but not caldesmon.

We isolated Ca(2+)-regulated thin filaments from the smooth muscle of the mussel Crenomytilus grayanus and studied the protein composition of different preparations from this muscle: whole muscle, heat-stable extract, fractions from heat-stable extract, thin filaments and intermediate stages of thin filaments purification. Among the protein components of the above-listed preparations, we did not find caldesmon (CaD), although two isoforms of a calponin-like (CaP-like) protein, which along with CaD is characteristic of vertebrate smooth muscle, were present in thin filaments. Thus, CaD is not Ca(2+)-regulator of thin filaments of this muscle. On the other hand, the mussel CaP-like protein is also not such Ca(2+)-regulator since we have shown that this protein can be selectively removed from isolated mussel thin filaments without loss of their Ca(2+)-sensitivity. We suggest that thin filaments in the smooth catch muscle possess other type of Ca(2+)-regulation, different from that in vertebrate smooth muscles.

Toxicity of cadmium and lead in Gallus gallus domesticus assessment of body weight and metal content in tissues after metal dietary supplements.

The influence of dietary cadmium on the accumulation and effects of dietary lead, examined in chicken. This experiment was conducted to investigate the toxic effects of dietary Cd and Pb on chick's body weight and organ, content of the tissues of these two metals was also detected. One day age chicks of Gallus gallus domesticus fed diet supplemented with 25, 50, 100 ppm of Cd, second group exposure to 300, 500, 1000 ppm of Pb in feed daily during 4 weeks. The control groups were fed without supplementation of metals. The concentrations of Cd and Pb resulted in increased of Cd and Pb content in liver, gizzard and muscle. While Cd 100 ppm and Pb 1000 ppm were increased metals content in feather. Body weight of chicks was not influenced by Cd treatment. In contrary Pb treatment was significantly (p < 0.05) decreased body weight of chicks after dietary treatment. On the other hand, Liver weigh in chicks was significantly (p < 0.05) decreased after Cd and Pb treatments.

The RNA-binding protein RBPMS2 regulates development of gastrointestinal smooth muscle.

BACKGROUND & AIMS: Gastrointestinal development requires regulated differentiation of visceral smooth muscle cells (SMCs) and their contractile activities; alterations in these processes might lead to gastrointestinal neuromuscular disorders. Gastrointestinal SMC development and remodeling involves post-transcriptional modification of messenger RNA. We investigated the function of the RNA-binding protein for multiple splicing 2 (RBPMS2) during normal development of visceral smooth muscle in chicken and expression of its transcript in human pathophysiological conditions. METHODS: We used avian replication-competent retroviral misexpression approaches to analyze the function of RBPMS2 in vivo and in primary cultures of chicken SMCs. We analyzed levels of RBPMS2 transcripts in colon samples from pediatric patients with Hirschsprung's disease and patients with chronic pseudo obstruction syndrome (CIPO) with megacystis. RESULTS: RBPMS2 was expressed strongly during the early stage of visceral SMC development and quickly down-regulated in differentiated and mature SMCs. Misexpression of RBPMS2 in differentiated visceral SMCs induced their dedifferentiation and reduced their contractility by up-regulating expression of Noggin, which reduced activity of bone morphogenetic protein. Visceral smooth muscles from pediatric patients with CIPO expressed high levels of RBPMS2 transcripts, compared with smooth muscle from patients without this disorder. CONCLUSIONS: Expression of RBPMS2 is present in visceral SMC precursors. Sustained expression of RBPMS2 inhibits the expression of markers of SMC differentiation by inhibiting bone morphogenetic protein activity, and stimulates SMC proliferation. RBPMS2 transcripts are up-regulated in patients with CIPO; alterations in RBPMS2 function might be involved in digestive motility disorders, particularly those characterized by the presence of muscular lesions (visceral myopathies).

Detection, accumulation, distribution, and depletion of furaltadone and nifursol residues in poultry muscle, liver, and gizzard.

Nitrofurans were broadly used as an extremely effective veterinary antibiotic especially in pig and poultry production farms. Because of fears of the carcinogenic effects on humans, the nitrofurans were banned from use in livestock production in many countries, including the European Union. The present study examines the accumulation, distribution, and depletion of furaltadone and nifursol and of their tissue-bound metabolites [3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) and 3,5-dinitro-salicylic acid hydrazine (DNSAH), respectively, in poultry edible tissues (muscle, liver, and gizzards) following administration to chickens of therapeutic and subtherapeutic concentrations of both compounds. Nitrofurans determination was performed by high-performance liquid chromatography-diode array detection and liquid chromatography-tandem mass spectrometry, respectively, for feeds and for poultry tissues. Furaltadone and nifursol, in very low concentrations, were found in samples of muscle, liver, and chicken's gizzard collected from slaughtered animals after 5 weeks of treatment and no withdrawal time period. When a withdrawal time period of 3 weeks was respected, no detectable nitrofuran parent compounds was observed in all of the studied matrices. For AMOZ, concentrations of 270 µg/kg in meat, 80 µg/kg in liver, and 331 µg/kg in gizzard were determined after administration of a medicated feed with furaltadone (132 mg/kg), 3 weeks after withdrawal of treatment. For DNSAH, the concentration values obtained are much lower than those observed for AMOZ. For meat, liver, and gizzard, DNSAH concentrations of 2.5, 6.4, and 10.3 µg/kg, respectively, were determined, after administration of a medicated feed with nifursol (98 mg/kg), 3 weeks after withdrawal of treatment. The gizzard could be considered a selected matrix for nitrofuran residues evaluation in poultry, due to its capacity of retaining either nitrofuran parent compounds or metabolites in higher concentrations, regardless of the administered dose or of the respected withdrawal time period.

Trace metal depositional patterns from an open pit mining activity as revealed by archived avian gizzard contents.

Archived samples of blue grouse (Dendragapus obscurus) gizzard contents, inclusive of grit, collected yearly between 1959 and 1970 were analyzed for cadmium, lead, zinc, and copper content. Approximately halfway through the 12-year sampling period, an open-pit copper mine began activities, then ceased operations 2 years later. Thus the archived samples provided a unique opportunity to determine if avian gizzard contents, inclusive of grit, could reveal patterns in the anthropogenic deposition of trace metals associated with mining activities. Gizzard concentrations of cadmium and copper strongly coincided with the onset of opening and the closing of the pit mining activity. Gizzard zinc and lead demonstrated significant among year variation; however, maximum concentrations did not correlate to mining activity. The archived gizzard contents did provide a useful tool for documenting trends in metal depositional patterns related to an anthropogenic activity. Further, blue grouse ingesting grit particles during the time of active mining activity would have been exposed to toxicologically significant levels of cadmium. Gizzard lead concentrations were also of toxicological significance but not related to mining activity. This type of "pulse" toxic metal exposure as a consequence of open-pit mining activity would not necessarily have been revealed through a "snap-shot" of soil, plant or avian tissue trace metal analysis post-mining activity.

The differences between the localizations of MUC1, MUC5AC, MUC6 and osteopontin in quail proventriculus and gizzard may be a reflection of functional differences of stomach parts.

Mucins are high molecular weight glycoproteins which constitute the major component of the mucus layer and are produce by many epithelial tissues in vertebrates. Osteopontin (OPN) is an adhesive phosphorylated glycoprotein that is expressed by a broad range of tissues and cells. Although gastric mucins MUC1, MUC5AC, MUC6 and OPN have been widely used in histological studies and in diagnostic pathology in order to diagnose gastric carcinomas, their localizations in the stomach of quail have not yet been studied. In this study, the localizations of MUC1, MUC5AC, MUC6 and OPN in the proventriculus and gizzard of Japanese quail during the post-hatching period were compared at light microscope levels by applying immunohistochemical methods. In all ages studied, the immunoreactivity of MUC5AC was present in the lining epithelium of both folds and superficial proventricular glands in the proventriculus, whereas MUC1, MUC6 and OPN reactivity was found in the oxynticopeptic cells of profound proventricular glands. In addition, some cells in the fold epithelium of the proventriculus showed a positive reaction to OPN. The immunoreactivity of MUC1 in gizzard was different from that of MUC5AC. Although MUC5AC was expressed in the cells of both the surface epithelium and profound glands of the gizzard, MUC1 was only localized in the profound glands of the gizzard. However, MUC6 and OPN immunoreactivity was absent in the gizzard. The results indicated that the differences between the localizations of MUC1, MUC5AC, MUC6 and OPN in quail proventriculus and gizzard may be a reflection of functional differences of stomach parts. Although the biological significances of the expressions of MUC1, MUC5AC, MUC6 and OPN in the quail stomach remains unknown, these notable glycoproteins may be associated with barrier function, host defence, and/or secretion.

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.

Avian digestive tract simulation to study the effect of grit geochemistry and food on Pb shot bioaccessibility.

Lead shot dissolution was investigated in a dynamic in vitro simulated avian gizzard-intestine system. The method allows simulated digestive fluid to pass (at intervals) from a gizzardlike environment to an intestine-based one, and then considers the dissolution of Pb shot (0-3 pellets) in the presence of differing grit geochemistries (siliceous and calcareous) and variable amounts of food (0-4 g of partially milled wheat seed). Dissolved Pb levels in simulated gizzards were consistently higher in the presence of siliceous, than with calcareous, grit. This was also seen in simulated intestines, except when less food was used (0-1 g), when Pb levels in solution were higher in calcareous systems. The Pb concentrations in gizzard and intestine solutions increased directly with the number of Pb shot used. In all treatments Pb levels in intestine liquids were lower than in gizzard liquids. Calcareous grit simulations maintained 2.5-34 times more Ca in solution than those that used siliceous grit. Dietary supplementation with calcareous grit may reduce Pb bioaccessibility of ingested Pb shot in birds by reducing gizzard acidity, by enhancing Pb precipitation (as Pb-carbonate), and by promoting higher dissolved Ca levels in the intestine, which may then compete with Pb for intestinal absorption.