Coccidiosis in Egg-Laying Hens and Potential Nutritional Strategies to Modulate Performance, Gut Health, and Immune Response.

Avian coccidiosis, despite advancements in management, nutrition, genetics, and immunology, still remains the most impactful disease, imposing substantial economic losses to the poultry industry. Coccidiosis may strike any avian species, and it may be mild to severe, depending on the pathogenicity of Eimeria spp. and the number of oocysts ingested by the bird. Unlike broilers, low emphasis has been given to laying hens. Coccidiosis in laying hens damages the gastrointestinal tract and causes physiological changes, including oxidative stress, immunosuppression, and inflammatory changes, leading to reduced feed intake and a drastic drop in egg production. Several countries around the world have large numbers of hens raised in cage-free/free-range facilities, and coccidiosis has already become one of the many problems that producers have to face in the future. However, limited research has been conducted on egg-laying hens, and our understanding of the physiological changes following coccidiosis in hens relies heavily on studies conducted on broilers. The aim of this review is to summarize the effect of coccidiosis in laying hens to an extent and correlate it with the physiological changes that occur in broilers following coccidiosis. Additionally, this review tries to explore the nutritional strategies successfully used in broilers to mitigate the negative effects of coccidiosis in improving the gut health and performance of broilers and if they can be used in laying hens.

Comparison of necrotic enteritis effects on growth performance and intestinal health in two different meat-type chicken strains Athens Canadian Random Bred and Cobb 500.

Chickens have undergone genetic improvements in the past few decades to maximize growth efficiency. However, necrotic enteritis (NE), an enteric disease primarily caused by C. perfringens, remains a significant problem in poultry production. A study investigated the differences in intestinal health between the nonselected meat-type chicken Athens Canadian Random Bred (ACRB) and the modern meat-type Cobb 500 broilers (Cobb) when challenged with experimental NE. The study utilized a 2 × 3 factorial arrangement, consisting of two main effects of chicken strain and NE challenge model (nonchallenged control, NC; NE challenge with 2,500/12,500 Eimeria maxima oocysts + 1 × 109C. perfringens, NE2.5/NE12.5). A total of 432 fourteen-day-old male ACRB and Cobb were used until 22 d (8 d postinoculation with E. maxima on d 14, dpi), and the chickens were euthanized on 6 and 8 dpi for the analysis. All data were statistically analyzed using a two-way ANOVA, and Student's t-test or Tukey's HSD test was applied when P < 0.05. The NE12.5 group showed significant decreases in growth performance and relative growth performance from d 14 to 20, regardless of chicken strain (P < 0.01). The ACRB group exhibited significant decreases in relative body weight and relative body weight gain compared to the Cobb group from d 14 to 22 (P < 0.01). On 6 and 8 dpi, both NE challenge groups showed significant decreases in intestinal villus height to crypt depth ratio, jejunal goblet cell count, and jejunal MUC2 and LEAP2 expression (P < 0.01). Additionally, the NE12.5 group had significantly higher intestinal NE lesion score, intestinal permeability, fecal E. maxima oocyst count, intestinal C. perfringens count, and jejunal IFNγ and CCL4 expression compared to the NC group (P < 0.05). In conclusion, NE negatively impacts growth performance and intestinal health in broilers, parameters regardless of the strain.

Effects of Methionine Supplementation Levels in Normal or Reduced Protein Diets on the Body Composition and Femur Bone Characteristics of Broilers Challenged with Coccidia.

This study investigated the effects of dietary methionine (Met) levels on the bone quality of broilers challenged with coccidia. A total of 600 fourteen-day-old male Cobb500 broilers were gavaged with mixed Eimeria spp. and randomly allocated into 10 treatment groups by a 2 × 5 factorial arrangement. Birds received normal protein diets (NCP) or reduced-protein diets (LCP), containing 2.8, 4.4, 6.0, 7.6, and 9.2 g/kg of Met. Data were analyzed via two-way ANOVA and orthogonal polynomial contrast. At 9 days post-inoculation (DPI), whole body bone mineral density (BMD) and bone mineral content (BMC) linearly decreased as Met levels increased (p < 0.05). For the femoral metaphysis bone quality at 9 DPI, BMD linearly decreased, and porosity linearly increased as Met levels increased (p < 0.05) in the cortical bone. The increased Met levels linearly improved trabecular bone quality in LCP groups (p < 0.05) while not in NCP groups. For the femoral diaphysis cortical bone at 6 DPI, LCP groups had higher BMD and BMC than NCP groups (p < 0.05). Bone volume linearly increased as Met levels increased in LCP groups (p < 0.05) while not in NCP groups. In summary, the results suggested that increased Met levels decreased the cortical bone quality. However, in the context of reduced-protein diets, the increased Met levels improved trabecular bone quality.

Aflatoxin B1 Impairs Bone Mineralization in Broiler Chickens.

Aflatoxin B1 (AFB1), a ubiquitous mycotoxin in corn-based animal feed, particularly in tropical regions, impairs liver function, induces oxidative stress and disrupts cellular pathways, potentially worsening bone health in modern broilers. A 19-day experiment was conducted to investigate the effects of feeding increasing levels of AFB1-contaminated feed (<2, 75-80, 150, 230-260 and 520-560 ppb) on bone mineralization markers in broilers (n = 360). While growth performance remained unaffected up to Day 19, significant reductions in tibial bone ash content were observed at levels exceeding 260 ppb. Micro-computed tomography results showed that AFB1 levels at 560 ppb significantly decreased trabecular bone mineral content and density, with a tendency for reduced connectivity density in femur metaphysis. Moreover, AFB1 above 230 ppb reduced the bone volume and tissue volume of the cortical bone of femur. Even at levels above 75 ppb, AFB1 exposure significantly downregulated the jejunal mRNA expressions of the vitamin D receptor and calcium and phosphorus transporters. It can be concluded that AFB1 at levels higher than 230 ppb negatively affects bone health by impairing bone mineralization via disruption of the vitamin D receptor and calcium and phosphorus homeostasis, potentially contributing to bone health issues in broilers.

Effects of phytase supplementation on broilers fed with calcium and phosphorus-reduced diets, challenged with Eimeria maxima and Eimeria acervulina: influence on growth performance, body composition, bone health, and intestinal integrity.

An experiment was conducted to evaluate the effects of phytase in calcium (Ca) and available phosphorous (avP)-reduced diet on growth performance, body composition, bone health, and intestinal integrity of broilers challenged with Eimeria maxima and Eimeria acervulina. A total of 672 14-day-old male broilers were allocated to a 2 × 4 factorial arrangement with 6 replicates per treatment and 14 birds per replicate. Two factors were Eimeria challenge and 4 dietary treatments: 1) a positive control (PC; 0.84% Ca and 0.42% avP); 2) a negative control (NC; 0.74% Ca and 0.27% avP); 3) NC + 500 FTU/Kg of phytase (NC + 500PHY); and 4) NC + 1,500 FTU/Kg of phytase (NC + 1500PHY). On d 14, birds in the Eimeria-challenged groups received a solution containing 15,000 sporulated oocysts of E. maxima and 75,000 sporulated oocysts of E. acervulina via oral gavage. At 5 d postinoculation (DPI), the challenged birds showed a higher (P < 0.01) FITC-d level than the unchallenged birds. While the permeability of the NC group did not differ from the PC group, the phytase supplementation groups (NC + 500PHY and NC + 1500PHY) showed lower (P < 0.05) serum FITC-d levels compared to the NC group. Interaction effects (P < 0.05) of Eimeria challenge and dietary treatments on feed intake (FI), mucin-2 (MUC2) gene expression, bone ash concentration, and mineral apposition rate (MAR) were observed. On 0 to 6 and 0 to 9 DPI, Eimeria challenge decreased (P < 0.01) body weight (BW), body weight gain (BWG), FI, bone mineral density (BMD), bone mineral content (BMC), bone area, fat free bone weight (FFBW), bone ash weight, bone ash percentage and bone ash concentration; and it showed a higher FCR (P < 0.01) compared to the unchallenged group. The reduction Ca and avP in the diet (NC) did not exert adverse effects on all parameters in birds, and supplementing phytase at levels of 500 or 1,500 FTU/Kg improved body composition, bone mineralization, and intestinal permeability, with the higher dose of 1,500 FTU/Kg showing more pronounced enhancements. There was an observed increase in FI (P < 0.01) when phytase was supplemented at 1,500 FTU/Kg during 0 to 6 DPI. In conclusion, results from the current study suggest that dietary nutrients, such as Ca and avP, can be moderately reduced with the supplementation of phytase, particularly in birds infected with Eimeria spp., which has the potential to save feed cost without compromising growth performance, bone health, and intestinal integrity of broilers.

Graded levels of Eimeria infection modulated gut physiology and temporarily ceased the egg production of laying hens at peak production.

An experiment was conducted to investigate the changes in gastrointestinal physiology, including intestinal leakage, immune response, oxidative stress, along with performance traits, of Hy-Line W-36 laying hens following Eimeria infection at peak egg production. A total of 360 laying hens, at 25 wk of age, were assigned randomly to 5 treatment groups, each consisting of 6 replicate cages, including a nonchallenged control group. The other 4 groups were inoculated with graded levels of mixed Eimeria species as high, medium-high, medium-low, and low doses, respectively. The body weight (BW) and body weight gain (BWG) of laying hens were measured from 0 to 14 days postinoculation (DPI). Average daily feed intake (ADFI) and hen-day egg production (HDEP) were measured from 0 to 15 and 0 to 28 DPI, respectively. Gut permeability was measured on 5 DPI, whereas oxidative stress, immune response, and expression of nutrient transporter genes were measured on 6 and 14 DPI. A significant linear reduction in BW and BWG was observed with increased Eimeria inoculation dosage on both 6 and 14 DPI (P < 0.001, P-Lin < 0.0001). An interaction between the Eimeria dosages and DPI was observed for ADFI (P < 0.0001). Feed intake in the challenged groups decreased starting at 4 DPI, with the most significant drop occurring at 7 DPI, which did not recover until 15 DPI. Following the challenge, gastrointestinal physiology shifted toward the host defense against the Eimeria infection by upregulating mRNA expression of tight junction proteins and immune responses while downregulating the expression of key nutrient transporters on 6 and 14 DPI (P < 0.05). An interaction between the Eimeria inoculation dosage and DPI was also observed for daily HDEP (P < 0.0001). Overall, HDEP was lower in the challenged groups compared to the control. Daily HDEP in the challenged groups dropped from 8 DPI and became similar to the control birds only after 24 DPI. Egg production temporarily ceased in most of the laying hens infected with the high and medium-high dosages of Eimeria. In conclusion, Eimeria infection activated the host immune response, negatively affecting the gastrointestinal health, growth performance, and temporarily ceased the egg production of Hy-Line W-36 laying hens when infected at peak production.

Osteoimmunology: A Link between Gastrointestinal Diseases and Skeletal Health in Chickens.

Bone serves as a multifunctional organ in avian species, giving structural integrity to the body, aiding locomotion and flight, regulating mineral homeostasis, and supplementing calcium for eggshell formation. Furthermore, immune cells originate and reside in the bone marrow, sharing a milieu with bone cells, indicating a potential interaction in functions. In avian species, the prevalence of gastrointestinal diseases can alter the growth and the immune response, which costs a great fortune to the poultry industry. Previous studies have shown that coccidiosis and necrotic enteritis can dramatically reduce bone quality as well. However, possible mechanisms on how bone quality is influenced by these disease conditions have not yet been completely understood, other than the reduced feed intake. On the other hand, several mediators of the immune response, such as chemokines and cytokines, play a vital role in the differentiation and activation of osteoclasts responsible for bone resorption and osteoblasts for bone formation. In the case of Eimeria spp./Clostridium perfringens coinfection, these mediators are upregulated. One possible mechanism for accelerated bone loss after gastrointestinal illnesses might be immune-mediated osteoclastogenesis via cytokines-RANKL-mediated pathways. This review article thus focuses on osteoimmunological pathways and the interaction between host immune responses and bone biology in gastrointestinal diseases like coccidiosis and necrotic enteritis affecting skeletal health.

Effects of Different Eimeria Inoculation Doses on Growth Performance, Daily Feed Intake, Gut Health, Gut Microbiota, Foot Pad Dermatitis, and Eimeria Gene Expression in Broilers Raised in Floor Pens for 35 Days.

The study was conducted to investigate the effects of different Eimeria inoculation doses on the growth performance, gut ecosystem, and body composition of broilers in floor pens for 35 days. A total of 750 15-day-old broilers were allocated to five experimental groups with six replicate pens. The five experimental groups included unchallenged control (CON); Eimeria dose 1 (ED1): E. acervulina: 31,250/E. maxima: 6250/E. tenella: 6250; Eimeria dose 2 (ED2): E. acervulina: 62,500/E. maxima: 12,500/E. tenella: 12,500; Eimeria dose 3 (ED3): E. acervulina: 125,000/E. maxima: 25,000/E. tenella: 25,000; and Eimeria dose 4 (ED4): E. acervulina: 250,000/E. maxima: 50,000/E. tenella: 50,000. On D 21, BW were linearly reduced by increased Eimeria inoculation doses (p < 0.01). On D 35, the Eimeria challenge groups had significantly lower BW compared to the CON group. Increased Eimeria inoculation doses linearly decreased crude fat (CF) (p < 0.01) on D 21. Increased Eimeria inoculation doses tended to increase the relative abundance of the phylum Proteobacteria (p = 0.098) on D 21. On D 35, lean:fat was linearly reduced by increased Eimeria inoculation doses (p < 0.05). Eimeria infection negatively influenced growth performance and gut health in broilers in the acute phase, and the negative effects were prolonged to D 35 in floor pen conditions.

Effect of Hydrogen Oxide-Induced Oxidative Stress on Bone Formation in the Early Embryonic Development Stage of Chicken.

The current study aimed to monitor the impact of H2O2-induced oxidative stress on avian bone formation during the early stage of embryonic development. Fertilized Cobb broiler eggs were divided into five treatment groups and micro-injected with varying concentrations of H2O2, i.e., control (PBS; 0 nM), 10 nM, 30 nM, 100 nM, and 300 nM, on embryonic day 3, with continued incubation thereafter. The treatment concentrations were selected based on the level of lipid peroxidation and the survival rate of embryo. Embryos were collected at 6 h, 24 h, 48 h, and 72 h post-injection. The mRNA expression levels of apoptotic markers, antioxidant enzymes, and early bone formation gene markers were measured. The results showed that the microinjection of H2O2 altered the expression pattern of antioxidant enzymes' mRNA during early embryogenesis and decreased the expression of COL1A2 and COL2A1 at 6 h and 24 h post-injection. Decreased expression of BMP, BGLAP, and RUNX2 was observed 48 h post-injection. Additionally, a shorter embryo length was observed in the 100 nM and 300 nM H2O2 treatment groups 72 h post-injection. In conclusion, H2O2-induced oxidative stress suppressed the expression of bone formation gene markers, with chronic effects on avian embryonic development.