Dietary AFETOXIN-BD supplementation mitigates multi-mycotoxin effects on performance, egg quality, and health status of late-laying hens

Abstract

Mycotoxins in poultry feed remain a major challenge to laying hen productivity, mainly in the late laying period when hens are at a higher risk of oxidative stress, compromised gut integrity and reduced reproductive performance. The efficacy of a commercial mycotoxin degrading preparation (AFETOXIN-BD) in reducing the negative impact of multi-mycotoxin contaminated feeds was evaluated. Two hundred 54-week-old Hy-Line Brown laying hens were randomly allotted to four dietary treatments (5 replicates of 10 hens) and fed for 35 days: a control diet (CON), a mycotoxin-contaminated diet (MT), CON plus 1,000 g/ton AFETOXIN-BD (BD), or MT plus 1,000 g/ton AFETOXIN-BD (MT+BD). The MT diet significantly (P < 0.05) reduced egg production rate, daily egg mass, eggshell strength, Haugh unit, antioxidant enzyme activities (T-SOD, GSH-Px, T-AOC), beneficial lactic acid bacteria populations and general health status of the hens, while increasing (P < 0.05) the feed-to-egg ratio, malondialdehyde (MDA) content, pathogenic bacteria counts (E. coli and Salmonella spp.), and the presence of mycotoxin residues in the eggs. Adverse effects were effectively mitigated (P < 0.05) in the MT+BD diet, while the BD diet exhibited better performance than the CON diet. Hens fed with BD diet showed (P < 0.05) superior production, egg quality, antioxidant activity, and intestinal microbial balance compared to all dietary treatments. From these results, it can be concluded that 0.1% AFETOXIN-BD supplementation is an effective nutritional intervention to mitigate the toxicity of multi-mycotoxin including AFB1, ZEN, FB1, OTA, and DON toxicity and improve productive efficiency, egg quality, and food safety in late-phase laying hens under commercial conditions, while also prolonging the productive lifespan.

Introduction

Mycotoxins are secondary toxic metabolites formed by fungi and are common in poultry diets around the world (Gurikar et al., 2023). In poultry diet, the most common and harmful mycotoxins are AFB1, deoxynivalenol (DON), zearalenone (ZEN), ochratoxin A (OTA) and fumonisin B1 (FB1). These mycotoxins can cause noteworthy poultry health, productivity, egg quality and food safety issues (Fakhri et al., 2025). Worldwide, around 25% of food crops are adulterated with mycotoxins each year, causing noteworthy economic losses (Eskola et al., 2020).

AFB1 is regarded a highly toxic aflatoxins (Yunus et al., 2011). The most detected mycotoxins are DON and its acetylated metabolites that can harm both human and bird health, causing symptoms such as abdominal pain, fever, nausea, headache, diarrhea and vomiting (Sobrova et al., 2010). Furthermore, ZEN has a similar structure to some natural estrogen. This property of ZEN is responsible for its binding to oestrogen receptors and causes anabolic and estrogenic effects in animals (Rogowska et al., 2019). Furthermore, AFB1, DON and ZEN have been reported to impair antioxidant capacity in poultry.

The late laying period (usually >50 weeks of age) is physiologically challenging for laying hens, involving a reduction in egg production persistency, egg quality, and immunity (Lee et al., 2025). During this period, mycotoxin exposure is further intensified by disrupting the reliability and purpose of the intestinal barrier, affecting nutrient uptake, oxidative stress, and the balance of reproductive hormones (Ma et al., 2025; Yuan et al., 2022; Zhao et al., 2021). Consequently, decreases in egg production, eggshell strength, Haugh unit, and flock health are often reported (Arulnathan et al., 2024).

Mycotoxin-degrading preparations have emerged as a practical and effective strategy to mitigate these adverse effects (Kihal et al., 2022). Some of these additives bind mycotoxins within the gastrointestinal tract, thus decreasing the bioavailability and the systemically toxic effects of mycotoxins, and some might also help maintain gut microbiota balance and antioxidant status (De Mil et al., 2017).

Given the heightened susceptibility of late-laying hens to mycotoxins, effective mitigation strategies are critically needed. This research aimed to compare the alleviating impact of a commercial mycotoxin degrading preparation in 54-week-old Hy-Line Brown laying hens fed a normal diet and a mouldy (mycotoxin contaminated) diet. Production performance, egg quality, antioxidant capacity, intestinal microbiota, egg contamination with mycotoxins, and health status were the important parameters that were evaluated. This research provides valuable insights for the poultry industry, demonstrating how targeted supplementation can extend productive life, improve egg quality and safety, and enhance economic outcomes in late-laying flocks.

Therefore, the current research hypothesized that the addition of 0.1% AFETOXIN-BD (a commercial mycotoxin degrading preparation) would effectively mitigate the adverse effects of multi-mycotoxin contaminated feed on overall performance, including production, egg quality, antioxidant status, intestinal microbiota balance, mycotoxin residues in eggs, and overall health in late-phase laying hens.

Materials and methods

Animal ethics statement

The experimental protocol was approved by the Animal Welfare Committee of Tangshan Finely Animal Care Co., Ltd., Tangshan, Hebei, China (reference no. TFAC-LH-2026-01). Birds were managed in accordance with the Guide for the Care and Use of Agricultural Animals in Research and Teaching (4th ed., 2020).

Experimental design and husbandry management

A total of 200 healthy 54-week-old late laying Hy-Line Brown hens with similar body weight, egg production rate, and health status were arbitrarily distributed into four dietary treatments, each of which had 5 replications of 10 hens. The experimental design is mentioned in Table 1.

Table 1.

Experimental design.

Treatment	CON	MT	BD	MT+BD
Diet	Control diet	Mycotoxin contaminated	Control diet + 1000 g/T AFETOXIN-BD	Mycotoxins + 1000 g/T AFETOXIN-BD
AFT (μg/kg)	0.32	4.56	0.32	4.56
ZEN (μg/kg)	15.23	268.48	15.23	268.48
DON (μg/kg)	47.19	858.74	47.19	858.74
OTA (μg/kg)	8.56	60.39	8.56	60.39
T-2 (μg/kg)	18.12	129.54	18.12	129.54
FB (μg/kg)	405.03	6986.28	405.03	6986.28

Mycotoxin contents in each treatment group were actual measured values.

Birds were kept in 3 tier battery cages. Each individual cage measures 40 cm in length and 45 cm in depth. Its front height is 45 cm, rear height 38 cm, with the base slope ranging from 6° to 8°. Feed and water were given freely. Feeding was done twice daily (at 08:00 and 15:00 h) and eggs were picked every day at 16:00 h. The three-day pre-experimental adaptation period was followed by 35-day experimental period.

Experimental diets

All experimental diets were formulated to meet the nutritional standards of laying hens in the late laying period (National Research Council, 1994). Four dietary treatments were used: a control diet (CON), a mycotoxin-contaminated diet (MT), CON supplemented with 1,000 g/ton AFETOXIN-BD (BD), and MT supplemented with 1,000 g/ton AFETOXIN-BD (MT+BD). The commercial mycotoxin-degrading preparation AFETOXIN-BD was manufactured by Tangshan Finely Animal Care Co., Ltd., China, and is composed of carefully selected probiotics and mycotoxin-degrading enzymes. The analyzed mycotoxin concentrations in the experimental diets are presented in Table 1, and the ingredient composition and nutrient levels of the basal diet are shown in Table 2.

Table 2.

The composition and nutrient levels of basal diets.

Ingredient (%)	Content	Nutritional Component	Content
Corn	50.63	Metabolizable energy (MJ/kg)	11.22
Soybean meal (43%)	17.4	Crude protein (%)	15.8
Corn DDGS	15	Crude ash (%)	11.85
Limestone Powder	9.81	Calcium (%)	3.48
Wheat bran	3.00	Total phosphorus (%)	0.60
Soybean oil	2.40	Lysine (%)	0.85
Calcium hydrogen phosphate	0.51	Methionine ± Cystine (%)	0.66
Sodium chloride	0.310
Choline chloride	0.2
L-Lysine sulfate (70%)	0.19
DL-Methionine (98.5%)	0.14
Premix*	0.41
Total	100

*The premix provided the following per kg of diet: Cu 20 mg, Fe 90 mg, Zn 90 mg, Mn 60 mg, I 0.65 mg, Se 0.35 mg, Vitamin A 9,000 IU, Vitamin D₃ 2,000 IU, Vitamin E 11 IU, Vitamin K 1.0 mg, Vitamin B₂ 5.8 mg, Niacin 66 mg, Pantothenic acid 10 mg, Vitamin B6 2.6 mg, Biotin 0.10 mg, Folic acid 0.7 mg, Vitamin B12 0.012 mg.

⁺Crude protein, crude ash, and calcium were measured values; metabolizable energy, total phosphorus, and amino acids were calculated values.

Production performance

The production performance parameters included daily egg collection, egg weight (EW), feed consumption, cracked eggs, and mortality were recorded. The residual feed was calculated on a weekly basis. Furthermore, egg production, average daily feed intake and egg weight, feed-to-egg ratio, cracked egg rate, and mortality rate were calculated using the following formulas (Ma et al., 2025).

Egg quality

At the end of each week, three eggs from each replicate were arbitrarily selected to assess egg quality. An egg analyser (EMT-7300, Robotmation Co., Ltd., Tokyo, Japan) was used to measure EW, albumen height (AH), Haugh unit (HU) and yolk color (YC). The strength of eggshells (ESS) was measured with a digital eggshell strength tester (AC220, Orka Company, Shanghai, China). The eggshell thickness without the inner membrane was taken at three points (blunt end, equator and sharp end) by an eggshell thickness gauge (Guilin Measuring Tool Cutting Tool Co., Ltd., Guilin, China) and then the mean value was recorded. Egg shape index, eggshell ratio (%) and yolk ratio (%) were measured as decribed by (Iqbal et al., 2025).

Antioxidant parameters

On the 35th day of the trial, five birds per treatment were randomly selected for antioxidant parameters. Approximately 5 mL blood samples were collected in anti-coagulant tubes from the wing vein, then centrifuged at 3000 rpm for 10 min, and the plasma was frozen at −20°C until it was analysed. The concentrations of total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px), catalase (CAT), malondialdehyde (MDA), and total antioxidant capacity (T-AOC) were measured in the plasma by using Nanjing Jiancheng Bioengineering Institute, Nanjing, China commercial kits (Hussain et al., 2025).

Intestinal microbiota composition

On d 35, one hen per replicate was randomly selected and euthanized by cervical dislocation, and the ceca were removed under sterile conditions. The digesta contents were diluted and plated on selective media to determine the microbial populations of Escherichia coli, Salmonella and lactic acid bacteria according to the protocol (Sedaghat and Torshizi, 2017).

Mycotoxin residues

To determine the presence of residues of mycotoxins in eggs, two eggs per replicate were arbitrarily collected at week 2. On the 35th day of the trial, the eggs were boiled for 15 minutes at 100°C. After cooling, the yolk and the egg white were separated and pooled for the determination of AFB1, DON, ZEA, OTA, and FB1 residues using LC-MS as performed by (Omar et al., 2023)

Health status

The general health status (consistency of faeces, condition of feathers, color of comb and hen behaviour) was noted and recorded daily.

Statistical analysis

Experimental data were analysed using one way ANOVA, using SPSS software (version 25.0, SPSS Inc., Chicago, IL, USA). Tukey’s post-hoc test was used for multiple comparisons. Data are given as mean ± SE (standard error). All differences were deemed to be statistically significant at a P < 0.05.

Results

Production performance

Production performance was lower in the MT diet compared with the other diets (P < 0.05) as shown in Table 3. Egg production rate and daily EM were significantly reduced, and feed to egg ratio was significantly increased in the MT group (P < 0.05). The production performance of both the BD and MT+BD diets was improved by supplementation with 0.1% AFETOXIN-BD. The overall performance of the BD diet was optimal with the highest EPR and EM and the lowest mortality rate when compared with the other treatments (P < 0.05).

Table 3.

Effect of AFETOXIN-BD on production performance during late laying period.

Treatment	CON	MT	BD	MT+BD	P-Value
Egg production rate (%)	82.39 ± 3.84ª	80.54 ± 3.05ᵇ	83.03 ± 4.39ª	82.48 ± 2.96ᵃᵇ	0.043
Daily feed intake (g)	115.14 ± 6.18	115.02 ± 5.28	115.03 ± 6.04	115.22 ± 3.10	0.999
Daily egg mass (g)	52.30 ± 4.16ª	50.61 ± 4.11ᵇ	52.78 ± 3.64ª	52.33 ± 2.95ª	0.038
Feed-to-egg ratio	2.20 ± 0.23ᵇ	2.28 ± 0.16ª	2.18 ± 0.11ᵇ	2.20 ± 0.16ᵇ	0.017
Average egg weight (g)	63.48 ± 4.47	62.84 ± 2.08	63.57 ± 3.73	63.44 ± 4.08	0.976
Cracked egg rate (%)	0.82 ± 0.02	0.85 ± 0.03	0.79 ± 0.03	0.81 ± 0.02	0.133
Mortality rate (%)	1.18 ± 0.12	2.47 ± 0.16	0.64 ± 0.02	1.01 ± 0.09	0.001

Values are reported as mean ± standard error (SE). Means in the same row having different super-scripts (a,b,c) are significantly different (P < 0.05).

Dietary groups were abbreviated as control diet (CON), mycotoxin contaminated diet (MT), control diet + 1000 g/T AFETOXIN-BD (BD), Mycotoxins + 1000 g/T AFETOXIN-BD (MT+BD).

Egg quality

The egg quality factors were significantly influenced by the MT diet (P < 0.05), as shown in Table 4. Significant differences were shown in both HU and ESS between the MT group and the other treatments (P < 0.05). Both groups of animals (BD and MT+BD) supplemented with AFETOXIN-BD showed a significant improvement in HU and ESS (P < 0.05). Furthermore, the HU, ESS, and AH of the eggs in the BD group were better compared with the CON group (P < 0.05).

Table 4.

Effect of AFETOXIN-BD on egg quality during late laying period.

Treatment	CON	MT	BD	MT+BD	P-Value
Eggshell thickness (mm)	0.33 ± 0.02	0.32 ± 0.02	0.34 ± 0.01	0.33 ± 0.01	0.079
Eggshell strength (N)	4.06 ± 0.32ᵇ	3.82 ± 0.32ᶜ	4.30 ± 0.30ª	4.25 ± 0.24ª	<0.001
Haugh unit	80.04 ± 0.61ᵇ	78.15 ± 0.52ᶜ	84.19 ± 0.53ª	83.38 ± 0.36ª	<0.001
Yolk color	6.90 ± 0.06	6.80 ± 0.03	7.00 ± 0.12	6.90 ± 0.11	0.065
Albumen height (mm)	6.92 ± 0.29ᵇ	6.85 ± 0.32ᵇ	7.11 ± 0.36ª	7.03 ± 0.41ᵇ	0.019

Values are reported as mean ± standard error (SE). Means in the same row having different super-scripts (a,b,c) are significantly different (P < 0.05).

Dietary groups were abbreviated as control diet (CON), mycotoxin contaminated diet (MT), control diet + 1000 g/T AFETOXIN-BD (BD), Mycotoxins + 1000 g/T AFETOXIN-BD (MT+BD).

Antioxidant

The MT diet affected the antioxidant status of hens as shown in Table 5. It markedly reduced the activities of T-SOD, GSH-Px and T-AOC and increased the concentration of MDA as compared with the other treatments (P < 0.05). Both BD and MT+BD groups exhibited significant improvement in T-SOD activity, GSH-Px activity and reduction in MDA level with supplementation of 0.1% AFETOXIN-BD (P < 0.001). The BD group had the highest antioxidant activity (P < 0.05).

Table 5.

Effect of AFETOXIN-BD on antioxidant capacity during late laying period.

Treatment	CON	MT	BD	MT+BD	P-Value
T-SOD (U/mL)	252.00 ± 3.28c	226.00 ± 6.65ᵇ	283.00 ± 5.62 a	276.00 ± 2.10ᵃ	<0.001
GSH-Px (U/mL)	2412.00 ± 100.29ᵇ	2368.21 ± 89.64ᵇ	2806.00 ± 96.53ᵃ	2720.00 ± 115.32ᵃ	<0.001
CAT (U/mL)	3.77 ± 0.43	3.64 ± 0.31	3.92 ± 0.37	3.84 ± 0.37	0.661
MDA (nmol/mL)	6.36 ± 0.09ᵇ	6.82 ± 0.22ᵃ	5.62 ± 0.12ᵇᶜ	5.78 ± 0.13ᵇᶜ	<0.001
T-AOC (U/mL)	7.88 ± 0.20ᵃᵇ	6.54 ± 0.16ᵇ	8.89 ± 0.31ᵃ	8.53 ± 0.22ᵃ	<0.001

Values are reported as mean ± standard error (SE). Means in the same row having different super-scripts (a,b,c) are significantly different (P < 0.05).

Dietary groups were abbreviated as control diet (CON), mycotoxin contaminated diet (MT), control diet + 1000 g/T AFETOXIN-BD (BD), Mycotoxins + 1000 g/T AFETOXIN-BD (MT+BD).

Intestinal microbiota

The results presented in Table 6 indicated that the MT diet significantly raised the counts of pathogenic bacteria (E. coli and Salmonella) and lowered the counts of beneficial lactic acid bacteria in the cecal digesta (P < 0.001). The supplementation of 0.1% AFETOXIN-BD was found to significantly improve the intestinal microbiota balance by decreasing E. coli and Salmonella counts and increasing the counts of lactic acid bacteria (P < 0.001) in both the CON and MT diets. The BD group had the best microbial profile.

Table 6.

Effect of AFETOXIN-BD on intestinal microbiota counts during late laying period.

Treatment	CON	MT	BD	MT+BD	P-Value
Escherichia coli (log 10 CFU/g)	7.12 ± 0.09ᵇ	8.26 ± 0.12ᵃ	6.43 ± 0.28ᶜ	6.78 ± 0.21ᶜ	<0.001
Salmonella (log 10 CFU/g)	5.98 ± 0.42ᵇ	6.53 ± 0.20ᵃ	5.30 ± 0.26ᶜ	5.59 ± 0.34ᶜ	<0.001
Lactic acid bacteria (log 10 CFU/g)	6.51 ± 0.16ᵇ	5.28 ± 0.22ᶜ	7.15 ± 0.23ᵃ	6.92 ± 0.32ᵃ	<0.001

Values are reported as mean ± standard error (SE). Means in the same row having different super-scripts (a,b,c) are significantly different (P < 0.05).

Dietary groups were abbreviated as control diet (CON), mycotoxin contaminated diet (MT), control diet + 1000 g/T AFETOXIN-BD (BD), Mycotoxins + 1000 g/T AFETOXIN-BD (MT+BD).

Mycotoxin residues in eggs

Table 7 shows that the MT diet produced the highest egg mycotoxin residues (AFB₁, DON, ZEN and OTA). AFETOXIN-BD supplementation significantly decreased the residues in both the BD and MT+BD groups. The lowest residues were found in the BD and MT+BD groups, and MT+BD also exhibited a significant decrease in residues compared with the MT diet.

Table 7.

Effect of AFETOXIN-BD on mycotoxins residues during late laying period.

Treatment	CON	MT	BD	MT+BD
Aflatoxin B₁ (ng/g)	1.11	6.06	0.32	3.92
Deoxynivalenol (ng/g)	3.51	15.94	0.62	9.25
Zearalenone (ng/g)	1.11	7.53	0.23	3.09
Ochratoxin (ng/g)	1.08	7.39	0.35	4.12

Residues in the MT group were the highest. The CON group showed the lowest residues among all treatments.".

Dietary groups were abbreviated as control diet (CON), mycotoxin contaminated diet (MT), control diet + 1000 g/T AFETOXIN-BD (BD), Mycotoxins + 1000 g/T AFETOXIN-BD (MT+BD).

Values are means of 5 replicates. Residues were highest in MT and lowest in BD.

Health status

Observations of health status made daily throughout the experimental period showed that the hens receiving the MT diet were in the worst condition regarding their health status, which included loose stools, passage of feed, pale combs, and poor feathering. The healthy overall condition of the birds in the BD group was indicated by normal faecal consistency, feed consumption, ruddy combs, and glossy feathers.

Discussion

The present study illustrates that multi-mycotoxin contamination (AFB1, ZEN, FB1, OTA, and DON) of late-phase laying hen diets causes systemic toxicity, as manifested by reduced production performance, poor egg quality, oxidative stress, gut dysbiosis, egg contamination with mycotoxins, and health deterioration. The adverse effects were mitigated through dietary supplementation with a clay mineral plus yeast cell wall extract (AFETOXIN-BD). Interestingly, the binder-supplemented diet without mycotoxin challenge was superior in several parameters, which indicates that there may be direct prebiotic and antioxidant effects in addition to the adsorption of toxins.

Mycotoxin challenge caused a significant reduction in egg production rate and EM, and a significant increase in feed: egg ratio (Zhao et al., 2021). These effects are mechanistically related to hepatorenal toxicity, impaired protein synthesis in the liver, impaired energy metabolism, and endocrine interference, which involves the effect of ZEN as a xenoestrogen on the hypothalamic-pituitary-gonadal axis. Susceptibility is increased by age-related ovarian decline in late-phase hens (Siloto et al., 2011; Yuan et al., 2022). AFETOXIN-BD restored performance parameters to levels comparable to or higher than the control, in large part due to its adsorptive properties: aluminosilicates bind polar toxins through cation exchange, while yeast β-glucans bind non-polar mycotoxins (ZEN and DON), which lowers their bioavailability. This is coherent with the results of Zhao et al. (2021) and a recent meta-analysis indicating that egg production increased by 4.2% and EW by 1.37 g when hens were fed the yeast cell wall extracts under mycotoxin challenge (Weaver et al., 2024). The better digestibility and gut health in the BD group suggests that yeast-derived mannan-oligosaccharides improve nutrient digestibility and gut health, whether they bind toxins (Baek et al., 2024).

Mycotoxins negatively affected eggshell strength and Haugh units, possibly because of disturbed calcium and phosphorus metabolism, inflammation of the uterine shell glands, and decreased albumen protein synthesis (Wei et al., 2024). AFETOXIN-BD completely recovered these parameters, and the BD group had better values than the control group. This improvement could be due to decreased intestinal inflammation, improved calcium absorption, and decreased oxidative damage to the intestinal epithelium (Zhao et al., 2021). These improvements are beneficial for both internal egg production and marketability.

The activities of T-SOD, GSH-Px, and T-AOC were significantly reduced, and MDA was significantly increased because of mycotoxin challenge, suggesting severe oxidative stress and mitochondrial dysfunction. These modifications align with previously reported alterations in lipid peroxidation and glutathione redox systems in laying hens following Fusarium mycotoxins (Kulcsár et al., 2021, 2024). AFETOXIN-BD has successfully countered this oxidative imbalance by controlling the absorption of toxins. The antioxidant status of the unchallenged BD group is also improved, underscoring the AFETOXIN-BD's intrinsic ability to enhance endogenous antioxidant mechanisms.

The consumption of the mycotoxin-contaminated diet caused an increase in the quantity of pathogenic E. coli and Salmonella spp. and a decrease in that of lactic acid bacteria. AFETOXIN-BD was used to restore the microbiota by adsorbing toxins (preserving tight junction proteins like ZO-1 and claudin-1) and by prebiotic effects on the growth of Lactobacillus and Bifidobacterium (Yuan et al., 2022; Zhai et al., 2022).

The detectable residues of AFB1, DON, ZEN,and OTA detectable residues in eggs of mycotoxin challenged group supports systemic absorption and transfer to developing oocytes (Antonissen, 2025). AFETOXIN-BD significantly lowered the residue levels for all toxins, with the lowest residues being found in the BD group. This decrease is important in terms of food security and public health because it enables the possibility of more efficient use of water and alleviates scarcity (Fakhri et al., 2021; Heuel et al., 2023; Zhu et al., 2023).

The MT group of health status hens had loose stools, undigested feed passed, pale combs and poor feathering, all of which were clinical signs of maldigestion, anaemia, oxidative stress and chronic inflammation. The results showed notable differences in faecal consistency, comb colour, and feather condition AFETOXIN-BD led to healthier, normal looking faeces, ruddy combs, and shiny feathers which highlighting significant enhancements in gut integrity, nutrient utilization and overall welfare.

Conclusion

To conclude, the supplementation of 0.1% AFETOXIN-BD during the late phase of egg-laying was effective in mitigating the adverse effects of multi-mycotoxin contamination in laying hens by enhancing production performance, egg quality, antioxidant status, gut microbiota balance, and decreasing egg mycotoxin residues. The binder was also effective in the unchallenged poultry, suggesting that it also has prebiotic and antioxidant properties. The results support the use of AFETOXIN-BD as a useful tool to optimize productivity, egg safety and hen health in the late phase of laying.

Authors’ contributions

Luke Liu designed the study, provided experimental materials, analyzed the data, critically revised the manuscript, approved the final version, and agrees to be accountable for all aspects of the work. Yongqiang Wang formulated the diets, managed the hens, drafted the manuscript, approved the final version, and agrees to be accountable for all aspects of the work. Shuangping Wang performed the laboratory analyses, critically revised the manuscript, approved the final version, and agrees to be accountable for all aspects of the work. Alison Zhao contributed to study planning and data interpretation, critically revised the manuscript, approved the final version, and agrees to be accountable for all aspects of the work.

Disclosures

All authors are employed by Tangshan Finely Animal Care Co., Ltd., China, which manufactures AFETOXIN-BD. This employment may constitute a potential conflict of interest. Nevertheless, this research aimed to evaluate the efficacy of AFETOXIN-BD as a dietary mycotoxin binder to improve production performance, egg quality, antioxidant status, gut microbiota, and egg safety in late-phase laying hens.