A. P. McElroy

Multi-platform next-generation sequencing of the domestic Turkey (Meleagris gallopavo): Genome assembly and analysis

The combined application of next-generation sequencing platforms has provided an economical approach to unlocking the potential of the turkey genome.

Influence of limestone and phytase on broiler performance, gastrointestinal pH, and apparent ileal nutrient digestibility

An experiment was conducted to determine the influence of 2 levels of dietary Ca from limestone and 3 levels of phytase on broiler performance, bone ash, gastrointestinal pH, and apparent ileal digestibility (AID) of Ca, P, and amino acids. Cobb 500 broilers (n = 576) were allowed access to one of 6 corn-soy diets from 0 to 16 d. Experimental diets contained 1.03% or 0.64% Ca from limestone and 0.61% total P. Each diet was supplemented with 0, 500, or 5,000 FTU/kg of phytase to create a 2 × 3 factorial experiment. Broiler feed intake (FI) and BW gain were not affected by dietary Ca or phytase. Feed conversion ratio was improved (P < 0.05) as dietary phytase increased (1.36, 1.34, and 1.31, respectively). Tibia ash percent was reduced (P < 0.05) from 41.4 to 40.0% as dietary Ca decreased but increased with phytase addition (P < 0.05). Gizzard and ileal pH were reduced (P < 0.05) in broilers fed 0.64% Ca compared with broilers fed 1.03% Ca. Phytase at 5,000 FTU/kg increased (P < 0.05) pH in the gizzard, duodenum, jejunum, and ileum. Apparent ileal P digestibility was increased (P < 0.05) in broilers fed 0.64% Ca compared with broilers fed 1.03% Ca (0.68 vs. 0.73, respectively). Apparent ileal Ca digestibility was increased (P < 0.05) in broilers fed 1.03% Ca compared with broilers fed 0.64% Ca (0.67 vs. 0.53, respectively). Phytase improved AID of CP in broilers fed 1.0% Ca but did not have an effect on AID of CP in broilers fed 0.64% Ca, which resulted in a Ca × phytase interaction (P < 0.05). In conclusion, high dietary Ca increased pH in gizzard and ileum and interfered with the AID of P and CP. The interactions between Ca and phytase in the gastrointestinal tract are complex, and feeding phytase at doses above industry recommendations may allow for reduced-Ca diets while maintaining broiler performance, bone ash, and improving amino acid digestibility.

Immune responses to dietary β-glucan in broiler chicks during an Eimeria challenge

Escalating consumer concerns regarding pathogen resistance have placed the poultry industry under mounting pressure to eliminate the use of chemotherapeutic agents as feed additives. One possible alternative receiving increased attention is the use of immunomodulators such as β-glucan. A study was conducted to investigate the effects of a yeast-derived β-glucan (Auxoferm YGT) on broiler chick performance, lesion scores, and immune-related gene expression during a mixed Eimeria infection. Day-old chicks were fed diets containing 0, 0.02, or 0.1% YGT. On d 8 posthatch, one-half of the replicate pens were challenged with a mixed inoculum of Eimeria acervulina, Eimeria maxima, and Eimeria tenella. Measurements were taken and samples collected on d 4, 10, 14, and 21 posthatch. Dietary supplementation had no effect on performance or mortality. On d 14, 3 birds per pen (n = 24/treatment) were scored for intestinal coccidia lesions. Gross lesion severity was significantly reduced in birds supplemented with 0.1% YGT. On d 10, inducible nitric oxide synthase (iNOS) expression was downregulated in the jejunum of challenged birds fed 0.1% YGT. Expression of iNOS in the ileum was downregulated in the nonchallenged birds, but upregulated in the challenged birds fed 0.1% YGT on d 14. Interleukin (IL)-18 was upregulated in the jejunum of 0.1% YGT-treated birds. Interferon (IFN)-γ expression was decreased in challenged and nonchallenged birds fed 0.1% YGT. The IL-4 expression was downregulated in the nonchallenged birds with 0.1% YGT diet supplementation. The IL-13 and mucin-1 levels were also reduced due to β-glucan supplementation. Mucin-2 expression was increased in the nonchallenged birds, but decreased in the infected birds fed 0.1% YGT. These results suggest that although Auxoferm YGT at doses of 0.02 and 0.1% does not influence performance, it significantly reduces lesion severity and is capable of altering immune-related gene expression profiles, favoring an enhanced T helper type-1 cell response during coccidiosis.

Extra-phosphoric effects of superdoses of a novel microbial phytase

An experiment was conducted to evaluate the influence of a novel microbial phytase on broiler performance from d 0 to 42 and tibia ash at d 21. Male Cobb 500 broilers (n = 2,016) 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 NC plus 500 (NC+500), 1,000 (NC+1,000), or 1,500 (NC+1,500) U/kg of microbial phytase. Diets were fed in 3 phases from d 0 to 21, d 22 to 42, and d 43 to 49 to 32 birds/pen and 9 replicate pens/diet. From d 0 to 21, broilers fed the NC diet had decreased (P ≤ 0.05) BW gain and tibia ash compared with broilers fed all other diets, except tibia ash in birds fed PC+500. Phytase supplementation at 500, 1,000, or 1,500 U/kg to the NC improved (P ≤ 0.05) BW gain and tibia ash comparable with the PC. Feed conversion ratio (FCR) was improved (P ≤ 0.05) in broilers fed NC+1,500 compared with broilers fed all other diets. From d 0 to 49, growth performance was not influenced (P > 0.05) by diet. However, FCR was improved (P ≤ 0.05) in broilers fed 1,500 U/kg of microbial phytase compared with broilers fed the PC, PC+DCP, and NC. There were no differences in performance or tibia ash between broilers fed the PC or PC+DCP, which would indicate the PC diet was sufficient in Ca and P. Therefore, the improvements in FCR in the NC+1500 may be associated with mitigation of the antinutrient effects of phytate rather than improved P utilization.

Performance and immune responses to dietary β-glucan in broiler chicks

During the first week posthatch, the avian immune system is immature and inefficient at protecting chicks from invading pathogens. Among immunomodulators, beta-glucans are known as biological response modifiers due to their ability to activate the immune system. Current research suggests that beta-glucans may enhance avian immunity; however, very little is known about their influence on regulation of immune function. A study was performed to evaluate the effects of dietary beta-glucan on growth performance, immune organ weights, peripheral blood cell profiles, and immune-related gene expression in the intestine. One-day-old chicks were fed a diet containing 0, 0.02, or 0.1% yeast beta-glucan (n = 30/treatment). On d 7 and 14 posthatch, body and relative immune organ weights were measured and small intestinal sections were collected to evaluate gene expression by quantitative real-time PCR. Peripheral blood samples were also collected to determine heterophil:lymphocyte ratios. Supplementation of beta-glucan did not significantly affect BW gains, and no significant differences were observed among groups for relative immune organ weights or heterophil:lymphocyte ratios. Compared with controls, expression of interleukin (IL)-8 was downregulated in the beta-glucan-treated groups on d 7 and 14. On d 14, beta-glucan inclusion resulted in increased inducible nitric oxide synthase expression. Expression of IL-18 was upregulated on d 7 but reduced on d 14 due to beta-glucan supplementation. On d 7, interferon-gamma and IL-4 expression decreased in the beta-glucan-treated groups. However, on d 14, IL-4 expression was upregulated in the supplemented groups. Intestinal expression of IL-13 was also downregulated in the beta-glucan-treated birds on d 7. These results suggest that dietary inclusion of beta-glucans altered the cytokine-chemokine balance; however, it did not elicit a robust immune response in the absence of a challenge, resulting in no deleterious effects on performance.

Effects of dietary enzymes on performance and intestinal goblet cell number of broilers exposed to a live coccidia oocyst vaccine

An experiment was conducted to evaluate the effects of dietary enzymes on performance, tibia ash, and intestinal goblet cells of broilers administered a live coccidia oocyst vaccine (Coccivac B, Schering Plough, Kenilworth, NJ). Cobb 500 straight-run broilers were obtained and one-half of the chicks were sprayed with the live coccidia oocyst vaccine. Chicks were weighed and placed in battery brooders with respect to nonvaccinated or vaccinated group according to dietary treatment. The 8 dietary treatments were a positive control (0.90% Ca and 0.45% available P), a negative control (NC; 0.80% Ca and 0.35% available P), NC + phytase (PHY), NC + protease (PRO), NC + xylanase (XYL), NC + PHY+ PRO, NC + PHY + XYL, and NC + PHY + PRO + XYL. A diet × vaccination interaction (P > 0.05) was not observed for feed intake or BW gain. Feed conversion ratio was improved (P ≤ 0.05) in birds fed NC + PHY + XYL compared with NC. Vaccination reduced (P ≤ 0.05) feed intake and BW gain from d 0 to 18. Tibia ash was reduced (P ≤ 0.05) in the NC and PRO or XYL diets. Vaccination increased goblet cell numbers in the duodenum of birds fed XYL, whereas no differences were found in goblet cell numbers between nonvaccinated and vaccinated birds in other dietary treatments, which resulted in a diet × vaccination interaction (P ≤ 0.05). Protease decreased and NC + PHY+ PRO increased goblet cells in the jejunum at d 7, which resulted in a diet × vaccination interaction (P ≤ 0.05). At d 18, NC + PHY + XYL was the only diet in which vaccination decreased goblet cells in the jejunum, resulting in a diet × vaccination interaction (P ≤ 0.05). The data indicate that NC + PHY + XYL improved the feed conversion ratio in broilers fed corn-soybean meal diets. The vaccination × dietary enzyme interaction altered the number of goblet cells in the small intestine. Dietary enzyme supplementation did not alleviate reductions in growth performance associated with the use of a live coccidia oocyst vaccine.

Eimeria Species and Genetic Background Influence the Serum Protein Profile of Broilers with Coccidiosis

Background Coccidiosis is an intestinal disease caused by protozoal parasites of the genus Eimeria. Despite the advent of anti-coccidial drugs and vaccines, the disease continues to result in substantial annual economic losses to the poultry industry. There is still much unknown about the host response to infection and to date there are no reports of protein profiles in the blood of Eimeria-infected animals. The objective of this study was to evaluate the serum proteome of two genetic lines of broiler chickens after infection with one of three species of Eimeria. Methodology/Principal Findings Birds from lines A and B were either not infected or inoculated with sporulated oocysts from one of the three Eimeria strains at 15 d post-hatch. At 21 d (6 d post-infection), whole blood was collected and lesion scoring was performed. Serum was harvested and used for 2-dimensional gel electrophoresis. A total of 1,266 spots were quantitatively assessed by densitometry. Protein spots showing a significant effect of coccidia strain and/or broiler genetic line on density at P<0.05−0.01 (250 spots), P<0.01−0.001 (248 spots), and P<0.001 (314 spots) were excised and analyzed by matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry. Proteins were identified in 172 spots. A total of 46 different proteins were identified. Of the spots with a corresponding protein identification, 57 showed a main effect of coccidia infection and/or 2-way interaction of coccidia infection×broiler genetic line at P<0.001. Conclusions/Significance Several of the metabolic enzymes identified in this study are potential candidates for early diagnostic markers of E. acervulina infection including malate dehydrogenase 2, NADH dehydrogenase 1 alpha subcomplex 9, and an ATP synthase. These proteins were detected only in Line A birds that were inoculated with E. acervulina. Results from this study provide a basic framework for future research aimed at uncovering the complex biochemical mechanisms involved in host response to Eimeria infection and in identifying molecular targets for diagnostic screening and development of alternative preventative and therapeutic methods.

Effect of incubation temperature on nutrient transporters and small intestine morphology of broiler chickens

This study evaluated the effects of elevated incubation temperature on posthatch nutrient transporter gene expression, integrity of the intestinal epithelium, organ development, and performance in Ross 308 broiler chickens. Ross × Ross 308 fertile eggs (n = 900) were incubated at different eggshell temperatures during development. From embryonic day (ED) 1 to ED12, all eggs were incubated at 37.1°C, whereas from ED13 to ED21, the eggs were divided into 2 groups for incubation at 37.4°C (S) or 39.6°C (H). Performance characteristics were measured at day of hatch (DOH) and d 7, 14, 21, 30, and 42. Small intestine and residual yolk sacs were collected at DOH and d 2, 4, 6, and 10 and weighed individually. Intestinal samples from the duodenum, jejunum, and ileum were evaluated for mucosal morphology and relative nutrient transporter gene expression. No significant differences were found in performance or organ weights. The intestinal morphology results showed a temperature × age interaction in duodenum villus height (P = 0.02) and crypt depth (P = 0.05) and in ileum villus height-to-crypt depth ratios (P = 0.02). There was a main effect of temperature, resulting in deeper crypts (P = 0.02) in the jejunum of chicks incubated at H compared with S. In the nutrient gene expression evaluation, peptide transporter (PepT1) showed a temperature × age interaction. On DOH and d 2, 4, and 10, PepT1 expression was similar between chicks incubated at S and H. However, on d 6, chicks incubated at S had significantly higher expression of PepT1 than those incubated at H. This study presents the effects of elevated incubation temperature on small intestine morphology and relative expression of nutrient transporter mRNA in high-yield broiler chicks, which can be important for the availability of nutrients and distribution of energy.

The effect of dietary protein level on performance characteristics of coccidiosis vaccinated and nonvaccinated broilers following mixed-species Eimeria challenge

A series of experiments were conducted to investigate the effect of starter diet protein levels on the performance of broilers vaccinated with a commercially available live oocyst coccidiosis vaccine before subsequent challenge with a mixed-species Eimeria challenge. Data indicated that an increasing protein concentration in the starter diet improved broiler performance during coccidiosis vaccination. Prechallenge performance data indicated that vaccination could decrease BW and increase feed conversion ratio. The time period most important for the observed effects appeared to be between 13 and 17 d of age. This reduction in performance parameters of vaccinated broilers compared with nonvaccinated broilers was eliminated by the conclusion of the experiments (27 d) in the diet groups with higher protein. Vaccination was effective at generating protective immunity against Eimeria challenge, as evidenced by increased (P < 0.05) BW gain, improved feed conversion, reduced postchallenge mortality, and reduced lesion development in vaccinated broilers compared with nonvaccinated broilers. These observations support numerous other reports that confirm live oocyst vaccination can be used effectively as a preventive against avian coccidiosis in commercially reared broilers. More important, these findings suggest that reduced protein concentration of starter diets can lead to significant losses in broiler performance when using a vaccination program to prevent coccidiosis.

An antimicrobial peptide is downregulated in the small intestine of Eimeria maxima-infected chickens

Avian coccidiosis is a major disease of poultry caused by the intestinal protozoa Eimeria. Infection leads to reduced feed efficiency and BW gain, resulting in severe economic losses for the poultry industry. Aviagen line A and line B birds show a differential response to Eimeria infection, with line B birds exhibiting higher lesion scores and mortality. The objective of this study was to examine differential intestinal gene expression between 2-wk-old line A and B chicks in response to a challenge with Eimeria maxima. After challenge with 1 × 10(4) oocysts/chick, more than 40% of line A chicks had lesion scores of 0 to 1 (scale of 0 to 4), similar to control chicks. In contrast, all line B chicks challenged at this same dose had lesion scores of 2 to 4. Total RNA was extracted from the jejunum of control and challenged chicks from both lines A and B. Microarray analysis revealed that liver-expressed antimicrobial peptide 2 (LEAP-2), a component of the innate immune system, was downregulated 20-fold in line A challenged chicks with lesion scores of 2 to 4 compared with line A control chicks, and was downregulated 11- to 71-fold in line B challenged chicks with lesion scores of 2 to 4 compared with line B control chicks. Liver-expressed antimicrobial peptide 2 was downregulated less than 2-fold in line A challenged chicks with lesion scores of 1 compared with line A control chicks, indicating that these chicks were similar to control chicks in their expression level of LEAP-2. Other genes (cytochrome P450, heat shock protein 25, keratin 19, and amino acid transporter ASCT1) showed different patterns of over- or underexpression. The expression of LEAP-2 was verified using real-time PCR, revealing a correlation between lesion score and magnitude of LEAP-2 downregulation for both line A and line B chicks. Thus, LEAP-2 may serve as a useful marker for identification of chickens resistant to E. maxima infection and potentially other Eimeria spp.