Circadian regulation of calcium and phosphorus homeostasis during the oviposition cycle in laying hens.

Maintenance of calcium and phosphorus homeostasis in laying hens is crucial for preservation of skeletal integrity and eggshell quality, though physiological regulation of these systems is incompletely defined. To investigate changes in mineral and vitamin D3 homeostasis during the 24-h egg formation cycle, 32-wk-old commercial laying hens were sampled at 1, 3, 4, 6, 7, 8, 12, 15, 18, 21, 23, and 24 h post-oviposition (HPOP; n ≥ 4). Ovum location and egg calcification stage were recorded, and blood chemistry, plasma vitamin D3 metabolites, circulating parathyroid hormone (PTH), and expression of genes mediating uptake and utilization of calcium and phosphorus were evaluated. Elevated levels of renal 25-hydroxylase from 12 to 23 HPOP suggest this tissue might play a role in vitamin D3 25-hydroxylation during eggshell calcification. In shell gland, retinoid-x-receptor gamma upregulation between 6 and 8 HPOP followed by subsequently increased vitamin D receptor indicate that vitamin D3 signaling is important for eggshell calcification. Increased expression of PTH, calcitonin, and fibroblast growth factor 23 (FGF23) receptors in the shell gland between 18 and 24 HPOP suggest elevated sensitivity to these hormones toward the end of eggshell calcification. Shell gland sodium-calcium exchanger 1 was upregulated between 4 and 7 HPOP and plasma membrane calcium ATPase 1 increased throughout eggshell calcification, suggesting the primary calcium transporter may differ according to eggshell calcification stage. Expression in shell gland further indicated that bicarbonate synthesis precedes transport, where genes peaked at 6 to 7 and 12 to 18 HPOP, respectively. Inorganic phosphorus transporter 1 (PiT-1) expression peaked in kidney between 12 and 15 HPOP, likely to excrete excess circulating phosphorus, and in shell gland between 18 and 21 HPOP. Upregulation of FGF23 receptors and PiT-1 during late eggshell calcification suggest shell gland phosphorus uptake is important at this time. Together, these findings identified potentially novel hormonal pathways involved in calcium and phosphorus homeostasis along with associated circadian patterns in gene expression that can be used to devise strategies aimed at improving eggshell and skeletal strength in laying hens.

Informal nutrition symposium: leveraging the microbiome (and the metabolome) for poultry production.

Knowledge of gut microbiology of poultry has advanced from a limited ability to culture relatively few microbial species, to attempting to understand the complex interactions between the bird and its microbiome. The Informal Nutrition Symposium 2021 was intended to help poultry scientists to make sense of the implications of the vast amounts of information being generated by researchers. This paper represents a compilation of the talks given at the symposium by leading international researchers in this field. The symposium began with an overview of the historical developments in the field of intestinal microbiology and microbiome research in poultry. Next, the systemic effects of the microbiome on health in the context of the interplay between the intestinal microbiota and the immune system were presented. Because the microbiome and the host communicate and influence each other, the novel field of kinomics (the study of protein phosphorylation) as used in the study of the poultry microbiome was discussed. Protein phosphorylation is a rapid response to the complex of signals among the microbiome, intestinal lumen metabolites, and the host. Then, a description of why an understanding of the role of microbial endocrinology in poultry production can lead to new understanding of the mechanisms by which the gut microbiota and the host can interact in defined mechanisms that ultimately determine health, pathogenesis of infectious disease, and behavior was given. Finally, a view forward was presented underscoring the importance of understanding mechanisms in microbiomes in other organ systems and other species. Additionally, the importance of the development of new -omics platforms and data management tools to more completely understand host microbiomes was stressed.

Short and Medium Chain Fatty Acids and Their Derivatives as a Natural Strategy in the Control of Necrotic Enteritis and Microbial Homeostasis in Broiler Chickens.

The use of antibiotic growth promoters (AGPs) has historically been the most important prophylactic strategy for the control of Necrotic Enteritis (NE) caused by some Clostridium perfringens toxin types in poultry. During the last five decades, AGPs have also been supplemented in feed to improve body weight gain and feed efficiency as well as to modulate the microbiome (consisting of microbes and their genes both beneficial and potentially harmful) and reduce enteric pathogens, among other benefits. New regulatory requirements and consumer preferences have led to strong interest in natural alternatives to the AGPs for the prevention and control of illnesses caused by enteric pathogens. This interest is not just focused on the direct removal or inhibition of the causative microorganisms but also the improvement of intestinal health and homeostasis using a range of feed additives. A group of promising feed additives is short- and medium-chain fatty acids (SCFA and MCFA, respectively) and their derivatives. The use of SCFA and MCFA, including butyric, caproic, caprylic, capric, and lauric acids, has shown strong effects against NE in broilers both at experimental and commercial levels. These fatty acids also benefit intestinal health integrity and homeostasis. Other effects have also been documented, including increases in intestinal angiogenesis and gene expression of tight junctions. Chemical modifications to improve stability and point of release in the intestine have been shown to improve the efficacy of SCFA and MCFA and their derivatives. The aim of this review is to give an overview of SCFA, MCFA and their derivatives, as an alternative to replace AGPs to control the incidence and severity of NE in poultry.

Effects of heat stress on performance, blood chemistry, and hypothalamic and pituitary mRNA expression in broiler chickens.

This study was conducted to evaluate potential hormonal mechanisms associated with the stress response, thermoregulation, and metabolic changes of broiler chickens exposed to high environmental temperature. Nine hundred 1-day-old male broiler chicks (Ross 708) were placed in floor pens and raised to 24 d. At 24 d, chicks were randomly assigned to 1 of 2 treatments, heat stress (HS) or no HS, and allocated into battery cages in 8 batteries (10 birds per cage, 2 cages per battery). On day 31, blood was collected prior to HS and analyzed using an iSTAT analyzer. Half of the batteries were then moved into 2 rooms with an elevated ambient temperature (35°C) for 8 h. The remaining batteries stayed in the thermoneutral rooms with an ambient temperature of 22°C. Beginning at 5 h after the initiation of HS, blood was collected and analyzed using an iSTAT analyzer, birds were euthanized, and hypothalamus and pituitary samples were collected (16 birds per treatment), flash frozen, and stored at -80°C until RNA extraction. Reverse transcription-quantitative PCR was used to compare mRNA levels of key corticotropic and thyrotrophic genes in the hypothalamus and pituitary. Levels of mRNA for each target gene were normalized to PGK1 (pituitary) and GAPDH (hypothalamus) mRNA. Differences were determined using mixed model ANOVA. HS decreased (P < 0.05) feed intake, BW, bicarbonate, potassium, CO2, and triiodothyronine, while it increased mortality, glucose, pH, plasma thyroxine, and corticosterone. Expression of pituitary corticotropin-releasing hormone receptor 1 was downregulated (P < 0.001), while corticotropin-releasing hormone receptor 2 mRNA levels were higher (P = 0.001) in HS birds. HS increased expression of thyroid hormone receptor ß (P = 0.01) (2.8-fold) and thyroid stimulating hormone ß (P = 0.009) (1.4-fold). HS did not affect levels of mRNA of genes evaluated in the hypothalamus. Results showed that HS significantly affected both the thyrotropic and corticotropic axes. Understanding the role and regulation of these pathways during HS will allow researchers to better evaluate management strategies to combat HS.

Effect of early neonatal development and delayed feeding post-hatch on jejunal and ileal calcium and phosphorus transporter genes expression in broiler chickens.

Calcium (Ca) and phosphorus (P) are essential minerals involved in many biological processes including bone development and mineralization. Plasma concentration of both minerals is tightly regulated, and Ca and P homeostasis is maintained via intestinal absorption, bone storage and exchange, and renal reabsorption. In the current broiler production systems, chicks are deprived of food and water for up to 72 h due to uneven hatching, hatchery procedures, and transportation time to farms. Post-hatch (PH) feed delay results in lower body and organ weight, higher feed conversion ratio and mortality, and delayed PH growth and GIT development. Little is known about the effects of early neonatal development and delayed or immediate feeding PH on Ca and P transporters. Therefore, the aim of the present study was to characterize expression patterns of Ca and P transporter genes in small intestine during the first 2 wk PH in chickens fed immediately after hatch (FED) or subjected to 48 h delayed feeding (NOTFED). Expression of all Ca and P transporters in jejunum and ileum was significantly (P < 0.05) affected by age. Among Ca transporter genes, only mRNA expression of Calbidin D28k in jejunum and Ca sensing receptor (CaSR) in ileum were significantly (P < 0.05) affected by delay in feed access. For P transporter genes' expression, only P transporter type III (PIT1) mRNA was significantly affected by age, delay in feed access, and their interaction (P < 0.05). In summary, we have shown, for the first time, early developmental changes of Ca and P transporter genes in broiler chickens. Results suggest that an increase in gene expression of some of the transporters corresponds with the switch from yolk to high starch diet. Overall, our results can be helpful in better understanding of Ca and P homeostasis in broilers.

Effects of early neonatal development and delayed feeding immediately post-hatch on the hepatic lipogenic program in broiler chicks.

The embryo to neonate transition is a critical period of development that has significant impact on broiler production. During this time important genetic programs governing metabolism and growth are established. The goal of this work was to study the effects of early post-hatch (PH) development and the time of initiation of feeding on activation of the genetic program regulating hepatic lipogenesis. A comparison of liver total RNA samples at hatch and 7 days PH was performed using oligonucleotide-based (Affymetrix GeneChip®) chicken genome microarrays. During the first week PH there was significant up-regulation of key lipogenic genes including: ATP citrate lyase (ACL), malic enzyme (ME), fatty acid synthase (FAS), acetyl-CoA carboxylase alpha (ACCα), stearoyl-CoA desaturase-1 (SCD-1), sterol regulatory element binding protein-2 (SREBP-2) and thyroid hormone responsive spot 14α (Spot 14α) among others. These findings were confirmed using gene-specific RT-PCR assays. In a follow-up study, we investigated the effects of withholding feed for the first 48 h PH (delayed feeding, DF) on lipogenic gene expression through 8 days PH. Body weight gain was significantly depressed by DF. Plasma levels of the major metabolic hormones that regulate lipogenic gene expression (insulin, glucagon and T(3)) changed significantly during PH development, but were largely unaffected by DF. Plasma glucose was significantly lower in the DF group at 24h PH but recovered thereafter. In general, DF inhibited the up-regulation of lipogenic genes until feeding was initiated. Delayed up-regulation was also observed for the lipogenic transcription factor genes, SREBP-1, SREBP-2 and peroxisome proliferator-activated receptor gamma (PPARγ), but not for carbohydrate response element binding protein (ChREB) or liver X receptor (LXR). Our results offer additional insight into the transcriptional programming of hepatic lipogenesis in response to the transition from high fat (yolk) to high carbohydrate (feed) nutrition that occurs during early PH development.

Modifying broiler diets with phytase and vitamin D metabolite (25-OH D(3)): impact on phosphorus in litter, amended soils, and runoff.

Adding phytase and 25- hydroxycholecalciferol (25-OH D(3)) to broiler diets has been shown effective at reducing total P concentrations in broiler litter. This study was conducted to determine the impact of field application of broiler litter from modified diets on P solubility in litter-amended soils and P losses in runoff. Five broiler diets and their resulting litters were evaluated: a high P diet, a low P diet, each of those basal diets with phytase added, and a low P diet with phytase and 25-OH D(3) added. A field study was initiated at two sites with each of the five broiler litters and a commercial P fertilizer (triple superphosphate [TSP]) applied at the same total P rate (150 kg P ha(-1)) and a control where no P was applied. Soil P was monitored over time at two depths (0-5 cm and 0-15 cm) soils were collected in the spring and fall to perform rainfall simulation studies. Broiler litter or TSP application increased soil water-soluble P and Mehlich 3-P concentrations relative to the control, however there were no consistent differences detected between litter treatments. Results from the rainfall simulation experiments indicate that diet modification with phytase or 25-OH D(3) does not increase the potential for P losses in runoff from amended soils relative to traditional diets. Moreover, broiler diet modification to reduce excreted P could be a potentially effective method for reducing watershed scale P surpluses in areas of intensive broiler production, without raising concerns over soluble P losses from litter-amended soils.

Detection of Salmonella enterica serovar typhimurium by using a rapid, array-based immunosensor.

The multianalyte array biosensor (MAAB) is a rapid analysis instrument capable of detecting multiple analytes simultaneously. Rapid (15-min), single-analyte sandwich immunoassays were developed for the detection of Salmonella enterica serovar Typhimurium, with a detection limit of 8 x 10(4) CFU/ml; the limit of detection was improved 10-fold by lengthening the assay protocol to 1 h. S. enterica serovar Typhimurium was also detected in the following spiked foodstuffs, with minimal sample preparation: sausage, cantaloupe, whole liquid egg, alfalfa sprouts, and chicken carcass rinse. Cross-reactivity tests were performed with Escherichia coli and Campylobacter jejuni. To determine whether the MAAB has potential as a screening tool for the diagnosis of asymptomatic Salmonella infection of poultry, chicken excretal samples from a private, noncommercial farm and from university poultry facilities were tested. While the private farm excreta gave rise to signals significantly above the buffer blanks, none of the university samples tested positive for S. enterica serovar Typhimurium without spiking; dose-response curves of spiked excretal samples from university-raised poultry gave limits of detection of 8 x 10(3) CFU/g.

Phytate phosphorus hydrolysis as influenced by dietary calcium and micro-mineral source in broiler diets.

Phytate phosphorus (PP) hydrolysis by a 3-phytase was studied in vitro at pH 2.5 and 6.5 with either 0, 1.0, 4.0, or 9.0 g of Ca/kg diet, or 0, 1.0, 5.0, 7.5, or 10.0 g/kg diet of micro-mineral premix added as inorganic (IMM) or an equivalent level as micro-mineral-amino acid complexes (MAAC). Adding Ca or micro-minerals reduced (P < 0.05) PP hydrolysis at both pHs; however, the effect was greater at pH 6.5. An in vivo experiment was conducted in which broilers were fed one of six diets for 30 h. The experimental design was a factorial of three micro-mineral forms (0 added, IMM, and MAAC) and two Ca levels (0 or 5 g/kg). Adding Ca reduced (P < 0.05) PP disappearance and increased Ca apparent absorption. No micro-minerals effect (P > 0.05) was seen. Therefore, in poultry diets, it is Ca that inhibits PP hydrolysis and decreases P availability.