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Coffee is a renowned beverage derived from plants globally. In the coffee production process, coffee pulp is a by-product that is abundant in phenolic compounds. Therefore, this study aimed to assess the effect of coffee pulp extract (CPE) on growth performance, blood biochemistry, intestinal morphology, carcass characteristics, and meat quality of broiler chickens. A total of 160 one-day-old male Ross 308 broilers were randomly allocated to four treatments with five replicates and eight chicks per replicate in a completely randomized design. These four dietary treatments included a basal diet with CPE of 0 (Control), 250 (CPE250), 500 (CPE500), and 1,000 (CPE1000) mg/kg diet for 35 days. The results showed that the body weight (BW) of the CPE500 group was significantly higher (P < 0.01) compared to the other groups at 35 days. Moreover, CPE500 increased the average daily gain (ADG) (P = 0.004) and reduced the feed conversion ratio (FCR) (P = 0.008). No significant differences (P > 0.05) were observed in the blood biochemistry profile. In addition, the investigation on intestinal morphology showed that CPE supplementation enhanced villus height (VH) (P = 0.004), crypt depth (CD) (P < 0.05), and ratio of VH:CD (P < 0.05) in the duodenum. Dietary supplementation with CPE significantly increased the percentage of neck weight (P < 0.05) compared to the control groups. However, no significant effects of CPE supplementation were observed on the meat quality parameters of breast and thigh muscles, including pH, color, water-holding capacity, and tenderness (P > 0.05). A significant increase (P < 0.05) in thigh fat content was observed with CPE supplementation. In conclusion, CPE500 can improve the growth performance and intestinal morphology of broiler chickens despite the presence of antioxidants and anti-inflammatory agents. This suggests that coffee pulp biomass could potentially be used as an alternative feed additive from agricultural biomass in broiler production., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025. Published by Elsevier Inc.)

Coffee cherry pulp (CCP) is a by-product of coffee bean production. CCP contains amounts of phenolic compounds that are beneficial for animals. This study evaluated the impact of coffee cherry pulp extract (CCPE) supplementation on growth performance, meat quality, carcass characteristics, serum biochemistry, cecum microbial population, intestinal morphology, and immune and antioxidant responses of broilers. Five hundred 1-day-old Ross 308 chicks were randomly assigned to five groups: a basal diet control, a basal diet with antibiotic growth promoters at 0.25 g/kg, and groups supplemented with CCPE at 0.5, 1.0, and 2.0 g/kg diet over 35 days. The results showed that throughout the experimental period, the groups supplemented with CCPE improved their final weight, average daily gain, and feed conversion ratio ( p < 0.05). CCPE at 1.0 and 2.0 g/kg diet reduced the average daily feed intake ( p < 0.05). In addition, CCPE at 0.5 g/kg reduced levels of serum alanine transaminase and aspartate aminotransferase ( p < 0.05). Triglyceride levels were the lowest in CCPE 2.0 ( p < 0.05). In the group supplemented with CCPE at all levels, the high-density lipoprotein levels significantly increased ( p < 0.05). Drip loss in the breast at 24 and 48 h decreased ( p < 0.05). Additionally, live weight, defeathered weight, and carcass weight significantly increased ( p < 0.05). Furthermore, CCPE improved intestinal morphology, especially villus height and the villus height per crypt depth ratio ( p < 0.05). CCPE supplementation also reduced pathogenic bacteria, increased Lactobacillus spp. ( p < 0.05), and increased the expression of immune-related genes and antioxidant activity in the liver and intestines ( p < 0.05). Therefore, the use of CCPE as an alternative to antibiotics in broiler feed improved growth performance and health parameters in broilers. It provides a sustainable and environmentally friendly option for supplementary feed, contributing to more efficient poultry nutrition management.

Approximately 10,000 metric tons of broiler livers are yielded every year in Taiwan. However, due to unpleasant odor and health concern, these livers are typically discarded as waste in the slaughtering stream in most developed or developed countries. In alignment with global agrocycle policies, a biofunctional chicken-liver hydrolysate (CLH) has been developed. This study was to investigate the effects of CLHs on glucose homeostasis and complications in type II diabetes. Insulin resistance was induced in liver (FL83B) and muscle (C2C12) cells using 30 and 20 ng TNF-α/mL, respectively, resulting in decreased glucose uptake and lower expressions of IRβ, p-Akt/Akt, and p-GSK3/GSK. CLH supplementation significantly upregulated (p<0.05) glucose uptakes and these proteins. In db/db mice, CLH supplementation improved insulin resistance, as shown by OGTT assay, HOMA-IR value and serum glucose levels, while also reducing serum lipids and liver damage indices (p<0.05). Additionally, CLH ameliorated (p<0.05) decreased hindlimb-gastrocnemius weight, and liver lipid contents, oxidative stress (sera and liver) and inflammatory cytokines. Increased glycogen accumulation was visualized in PAS-stained liver and hindlimb tissues of db/db mice supplemented with CLHs, consistent with upregulated glycogenesis in TNF-α-induced liver and muscle cells through the IRβ-Akt-GSK3 pathway. These findings suggest CLH may offer a mitigation against hyperglycemia and associated complications in type II diabetes, while also highlighting a sustainable solution for utilizing poultry slaughter residues., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Background and Aim: The success of semen cryopreservation relies on several aspects, including breed, age, season, collection method, extender composition, cooling rate, equilibration period, freezing rate, and thawing rate. This study aimed to investigate the effects of cooling and equilibration duration, as well as the addition of antioxidants to the semen extender, on the cryopreservation of swamp buffalo semen., Materials and Methods: Semen collected from swamp buffalo bulls was subjected to four different conditions: (T1) 2-h cooling and 2-h equilibration, (T2) 1.5-h cooling and 1.5-h equilibration, (T3) 1-h cooling and 1-h equilibration, and (T4) 0.5-h cooling and 0.5-h equilibration. Spermatozoa motility was evaluated using a computer-assisted semen analyzer. Moreover, this study also investigated the effect of antioxidant supplementation during cryopreservation using tris-citrate egg yolk extenders enriched with various antioxidants: Control (Con), 1 mM melatonin (ML), 0.5 mM gamma-oryzanol (GO), 10 μM canthaxanthin (CX), 1 mM melatonin + 0.5 mM gamma-oryzanol (ML + GO), and 1 mM melatonin + 10 μM canthaxanthin (ML + CX)., Results: Results showed that the (T1) 2-h cooling and 2-h equilibration and (T2) 1.5-h cooling and 1.5-h equilibration groups achieved higher progressive motility than the (T3) 1-h cooling and 1-h equilibration and (T4) 0.5-h cooling and 0.5-h equilibration groups. The ML-treated group exhibited superior progressive motility and total motility., Conclusion: The optimal approach for cryopreserving swamp buffalo bull semen involves a 1.5-h cooling period followed by a 1.5-h equilibration period, with the incorporation of ML into the semen extender., Competing Interests: The authors declare that they have no competing interests., (Copyright: © Inyawilert, et al.)

The utilization of mangosteen biomass not only solves environmental problems but also raises the value of agricultural waste. The current study aimed to evaluate the potential of mangosteen pericarp extract (MPE) for enhancing the immunity and productivity of Thai native crossbred chickens on-farm. A total of 180 three-week-old chickens were divided into negative control and supplemented groups, with 1000 mg MPE/kg of diet. The safety of MPE was further confirmed by the absence of noticeable differences in mortality and biochemical parameters during the entire study period. The MPE-supplemented group displayed significant differences in the relative transcription levels of IL-10 compared to the basal diet group ( p ≤ 0.01). Preslaughter body weight, average daily gain, and carcass weight in the MPE-supplemented group were higher than those in the basal diet group ( p ≤ 0.05). Furthermore, MPE supplementation improved meat quality by enhancing the nutritional composition of protein and fat ( p ≤ 0.05), as well as improving water-holding capacity, lowering boiling, and lowering grilling losses ( p ≤ 0.01). These findings indicate that MPE can be an effective supplement for enhancing flock immunity, growth performance, and meat quality in poultry. This contributes to more sustainable agriculture and food security within agroecosystems.

Public interest in the cannabis plant has increased after its legalization in many countries. Cannabis sativa residues (CR) are a part of the plant waste in the cannabis industry. The CR contain medicinal properties that could be used as a feed additive in poultry production. The trial was conducted to investigate the effects of CR on growth performance, carcass characteristics, intestinal morphology, and blood biochemistry profile of broiler chickens. In a completely randomized design, 256 one-day-old male Ross 308 broilers were randomly allocated to 4 treatments with 8 replicates and 8 birds per replicate. These 4 dietary treatments included a basal diet with 0, 0.5, 1 and 2% CR for 40 d. The results showed that 2% CR supplementation reduced feed intake (FI) in the starter phase (d 3-23, P < 0.05). The birds in the CR groups had lower FI in the finishing phase (d 24-40, P < 0.01) and the whole raising period (d 3-40, P < 0.01) than the control. However, the body weight and carcass yield were not different (P > 0.05). In addition, the CR diet had no adverse effects on the blood biochemistry profile, including total cholesterol, triglycerides, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, total protein, globulin, albumin, and direct bilirubin (P > 0.05). In addition, total bilirubin and malondialdehyde were better in the plasma of CR-supplemented birds than in the control groups (P < 0.05). The observations on intestinal morphology showed that CR supplementation improved the ratio between villus height and crypt depth in the ileum (P < 0.05). In conclusion, CR supplementation can improve intestinal morphology and oxidative stability of broiler chickens. This suggests that CR could potentially be used as an alternative feed additive in broiler production., Competing Interests: DISCLOSURES The authors declare no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

This study aimed to assess the effectiveness of aflatoxin B 1 (AFB 1 ) and Thunbergia laurifolia extract (TLE) in the diets of Cherry Valley ducklings. Our investigation covered growth indicators, blood biochemical indices, meat quality, intestinal morphology, immune response, and CP450 enzyme-related gene expression. We conducted the study with 180 seven-day-old Cherry Valley ducks, randomly divided into five dietary treatments. These treatments included a basal diet without AFB 1 (T1 group), TLE, or a commercial binder; the basal diet containing 0.1 mg AFB 1 /kg (T2 group), 0.1 mg AFB 1 /kg and 100 mg TLE/kg (T3 group), 0.1 mg AFB 1 /kg and 200 mg TLE/kg (T4 group), and 0.1 mg AFB 1 /kg and 0.5 g/kg of a commercial binder (T5 group), respectively. Ducklings fed with the T2 diet exhibited lower final body weight (BW), average body weight gain (ADG), and poor feed conversion ratio (FCR) during the 42-day trials. However, all ducklings in the T3, T4, and T5 groups showed significant improvements in final BW, ADG, and FCR compared to the T2 group. Increased alanine transaminase (ALT) concentration and increased expression of CYP1A1 and CYP1A2 indicated hepatotoxicity in ducklings fed the T2 diet. In contrast, ducklings fed T3, T4, and T5 diets all showed a decrease in the expression of CYP1A1 and CYP1A2, but only the T4 treatment group showed improvement in ALT concentration. AFB 1 toxicity considerably raised the crypt depth (CD) in both the duodenum and jejunum of the T2 group, while the administration of 200 mg TLE/kg (T4) or a commercial binder (T5) effectively reduced this toxicity. Additionally, the villus width of the jejunum in the T2 treatment group decreased significantly, while all T3, T4, and T5 groups showed improvement in this regard. In summary, T. laurifolia extract can detoxify aflatoxicosis, leading to growth reduction and hepatic toxicosis in Cherry Valley ducklings.

The immunological sexing method using antibodies offers cost-effective, high-volume production but faces challenges in terms of X-sperm purity in sexed semen. This research aimed to produce sexed bull semen using highly specific recombinant antibodies in magnetic-activated cell sorting (MACS), evaluate sperm quality and kinematic parameters, and verify the sex ratio of sperm, embryos, and live calves. Fresh semen from two Angus bulls was separated into two equal groups: conventional (CONV) semen and semen sexed using MACS with Y-scFv antibody conjugation to separate two fractions, i.e., the X-enriched and Y-enriched fractions. Then, computer assisted semen analysis and imaging flow cytometry were used to evaluate sperm motility and kinematic variables, acrosomal integrity, sperm viability, and sperm sex ratios. The results showed that sperm motility and quality did not differ between X-enriched and CONV semen. However, the Y-enriched fraction showed significantly lower sperm quality than the X-enriched fraction and CONV semen. The sperm ratio revealed that X-sperm accounted for up to 79.50% of the X-enriched fraction, while Y-sperm accounted for up to 78.56% of the Y-enriched fraction. The sex ratio of embryos was examined using in vitro fertilization. The cleavage rates using CONV and X-enriched semen were significantly higher than that using Y-enriched semen. Accordingly, 88.26% female blastocysts were obtained by using X-enriched semen, and 83.58% male blastocysts were obtained by using Y-enriched semen. In farm trials, 304 cows were subjected to AI using X-enriched and CONV semen. The pregnancy rate did not differ between the X-enriched and CONV semen groups. On the other hand, X-enriched semen generated significantly more live female calves (83.64%) than CONV semen (47.00%). The MACS sexing method significantly enhanced the X-sperm purity in sexed semen, producing high-quality sperm, a high percentage of female blastocytes, and a high percentage of live female calves., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: At present, porcine reproductive and respiratory syndrome (PRRS) caused by the PRRS virus (PRRSV) is one of the most severe epidemics impacting pig farming globally. Despite the fact that a number of studies have been conducted on potential solutions to this problem, none have proven effective. The focus of problem solving is the use of natural ingredients such as plant extracts. Popular throughout Asia, Caesalpinia sappan (CS) is a therapeutic plant that inhibits PRRSV in vitro. Therefore, this study was performed to determine the efficacy of CS extract dietary supplementation on the productive performance, antibody levels, immunological indicators, and lung pathology of PRRSV-challenged weaned pigs. A total of 32 weaned piglets (28 days old) were randomized into 4 groups and kept separately for 14 days. The treatments were organized in a 2 × 2 factorial design involving two factors: PRRSV challenge and supplementation with 1 mg/kg CS extract. The pigs in the PRRSV-challenged groups were intranasally inoculated with 2 mL of PRRSV (VR2332) containing 10 4 TCID 50 /mL, while those in the groups not challenged with PRRSV were inoculated with 2 mL of normal saline., Results: In the PRRSV-challenged group (CS + PRRSV), supplementation with CS extract led to an increase in white blood cells (WBCs) on Day 7 post infection (p < 0.05) and particularly in lymphocytes on Days 7 and 14. The antibody titer was significantly greater in the CS + PRRSV group than in the PRRSV-challenged group not administered CS (PRRSV group) on Day 14 postinfection (S/P = 1.19 vs. 0.78). In addition, CS extract administration decreased the prevalence of pulmonary lesions, which were more prevalent in the PRRSV-challenged pigs that did not receive the CS extract., Conclusion: The findings of this study suggest that supplementation with CS extract is beneficial for increasing WBC counts, especially lymphocytes, increasing the levels of antibodies and reducing the prevalence of lung lesions in PRRSV-infected pigs., (© 2024. The Author(s).)

The present study investigated the effects of dietary supplementation of Caesalpinia sappan Linn Extract (CSE) on the health and productive performance of late-phase laying hens on farms. Proximate composition and antioxidant markers of CSE powder revealed favorable characteristics with high total dry matter; phenolic content, and antioxidant potency. Three hundred and sixty (64-week-old) Hy-line Brown hens were divided into five groups with 0 (control diet), 250, 500, 1000, and 2000 mg/kg CSE, respectively. The laying performance and egg quality of the CSE supplementation groups demonstrated significant improvements in egg weight and albumin weight ( p < 0.05), and a tendency for enhanced egg mass and feed conversion ratio. Additionally, the intestinal morphostructural indices in the 2000 mg CSE/kg diet group showed the greatest statistical significance ( p < 0.05), with a detectable trend suggesting an increase in the villus height to crypt depth ratio. In addition, significant downregulation of proinflammatory genes occurred in their liver tissues, coupled with a greater expression of genes linked to antioxidants and anti-inflammatory processes. Furthermore, the blood biochemical parameters and the organ weights may suggest a favorable safety profile of CSE supplementation. These findings highlight the potential of CSE as a dietary supplement to enhance the productive performance and flock health of late-phase laying hens. Further research is warranted to explore the long-term effects and optimal dosage of CSE supplementation for laying hens in farming practices.

Immune-mediated hemolytic anemia (IMHA) is a common autoimmune disorder in dogs with a high fatality rate and it remains a therapeutic challenge. The marine lipid extract, EAB-277, is a natural anti-inflammatory nutraceutical product. However, the effects of EAB-277 in IMHA dogs has rarely been investigated. The objective of this study is to assess the clinical effects of EAB-277 and prednisolone dose-tapering for supplemental therapy in IMHA dogs. Prednisolone was given to 18 anemic IMHA dogs according to a standard regimen. Six dogs were supplementally treated with EAB-277 for 28 days and the remaining twelve dogs were a control group of untreated supplementations. The results demonstrate that the supplement group showed slightly better survival rates (66.7 ± 19.2%) than the control group (16.7 ± 0.7%), but the difference was not statistically significant ( p = 0.408). When compared to pre-therapy, the supplement group's blood profiles improved ( p < 0.05). The EAB-277 treated group showed a moderate decrease in the incidence rate (4.20 times) of prednisolone tapering compared to the control group. The dosage reduction of prednisolone in supplement group was more than that in the control group ( p < 0.0001). Our results suggest that EAB-277 supplementation may enhance clinical outcomes and lessen prednisolone dose-tapering in canine IMHA therapy.

This research aimed to identify bioactive compounds from Caesalpinia sappan extract that function as novel porcine reproductive and respiratory syndrome virus (PRRSV) infection inhibitors by computational molecular screening. We obtained a set of small-molecule compounds predicted to target the scavenger receptor cysteine-rich domain 5 (SRCR5) of CD163. In addition, the functions of positive hits were assessed and verified utilizing an in vitro antiviral activity assay with PRRSV-infected MARC-145 cells. Combining molecular docking with the results of binding affinity and ligand conformation, it was found that brazilin had the highest binding energy with the SRCR5 receptor compared to catechin and epicatechin (- 5.8, - 5.5, and - 5.1 kcal/mol, respectively). In terms of molecular mechanics, the binding free energy between the SRCR5 receptor was - 15.71 kcal/mol based on the Poisson-Boltzmann surface area of brazilin. In addition, PRRSV infection in MARC-145 cells was significantly inhibited by brazilin compared to the control (virus titer, 4.10 vs. 9.25 TCID 50 /mL, respectively). Moreover, brazilin successfully limited the number of PRRSV RNA copies in MARC-145 cells as determined by RT-qPCR. By inhibiting the PRRSV-CD163 interaction with brazilin from Caesalpinia sappan, it may be possible to prevent PRRSV infection in pigs, as suggested by this research., (© 2022. The Author(s).)

This study aimed to investigate the effects of mango peel powder (MGPP) on growth, innate immunity, and immune-antioxidant related gene expression of Nile tilapia reared under biofloc system. Three hundred Nile tilapia (average weight 14.78 ± 0.05 g) were distributed into 15 fiber tanks (300 L per tank) assigned to five treatments in triplication. Fish were fed basal diet containing different levels MGPP as follows: 0 (MGPP0: control), 6.25 (MGPP 6.25), 12.5 (MGPP 12.25), 25 (MGPP 25), and 50 (MGPP 50) g kg -1 diet for 8 weeks. Specific growth rate (SGR), weight gain (WG), final weight (FW), feed conversion ratio (FCR), skin mucus of lysozyme (SMLA), and peroxidase activities (SMPA), serum of lysozyme (SL) and peroxidase (SP) were measured every for weeks; while immune-antioxidant-related gene expressions were determined after 8 weeks post-feeding. The results indicated that MGPP 25 diet resulted in higher SGR, WG, FW, and FCR but no significant differences among treatments were noticed. In terms of immune responses, lysozyme and peroxidase activities in mucus and serum were significantly higher in MGPP 12.5 and MGPP 25 diets against the control. Similarly, significant up-regulation of IL-1 and IL-8 gene expressions was observed in fish fed MGPP 25 against the control. However, no significant differences in LBP, GSTa, GPX, and GSR among treatments were observed. Overall, dietary inclusion of MGPP 25 significantly enhanced immune response and immune related gene expressions but not growth performance and antioxidant gene expressions. The results implied that MGPP can be potentially used as an immunostimulants in Nile tilapia culture., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Mycotoxin-contaminated feeds may negatively affect broiler chickens’ health; hence, a sustainable approach to achieve mycotoxin elimination is necessary. This study aimed to evaluate the efficacy of red yeast (Sporidiobolus pararoseus; RY) as a novel mycotoxin binder in broilers. A total of 1440 one-week-old male broiler chicks were randomly assigned to 12 treatments in a 3 × 4 factorial design. The dietary treatments included three levels of mycotoxin-contaminated diets (0 µg kg−1 (0% of mycotoxin; MT), 50 µg kg−1 (50% MT), and 100 µg kg−1 (100% MT)) and four levels of mycotoxin binders (0.0 and 0.5 g kg−1 commercial binder, and 0.5 and 1.0 g kg−1 RY). Experimental diets were contaminated with aflatoxin B1, zearalenone, ochratoxin A, T-2 toxin, and deoxynivalenol in the basal diet. Furthermore, the parameters including feed intake, body weight, and mortality rate were recorded on a weekly basis. After feeding for 28 days, blood and organ samples were collected randomly to determine the blood biochemistry, relative organ weights, and gut health. The results indicated that mycotoxin-contaminated diets reduced the average daily weight gain (ADG), villus height (VH), and villus height per the crypt depth ratio (VH:CD) of the intestine, as well as the population of Lactobacillus sp. and Bifidobacterium sp. in the cecal (p < 0.05), whereas they increased the mycotoxins concentration in the blood samples and the apoptosis cells (TUNEL positive) in the liver tissue (p < 0.01) of broiler chicken. In contrast, RY-supplemented diets had better ADG values and lower chicken mortality rates (p < 0.05). Moreover, these combinations positively impacted the relative organ weights, blood parameters, bacteria population, intestinal morphology, and pathological changes in the hepatocytes (p < 0.05). In conclusion, RY supplementation effectively alleviated the toxicity that is induced by AFB1 and OTA, mainly, and could potentially be applied as a novel feed additive in the broiler industry.

Relatively, little is known about the corpus luteum (CL) function in early pregnancy after the successful treatment of luteal phase deficiency in repeat-breeder dairy cows when exposed to extreme environments under tropical climate. To investigate the influence of increased tissues of corpora lutea (CLs) by inducing secondary CL based on progesterone (P4) concentration and fertility in repeat-breeder dairy cows undergoing the fixed-time artificial insemination (FTAI) protocol, 32 cows were treated with gonadotropin releasing hormone (GnRH) on day 5 post-induction (experiment 1). In experiment 2, 213 cows were bred using the short-term FTAI protocol. On day 5 post-FTAI, cows were divided into two groups: treatment with (GnRH5-treated group) or without (GnRH5-untreated group) GnRH. The temperature-humidity index ranged from 77.3 to 82.8. Cows bearing two CLs had greater P4 concentrations than cows bearing only one CL on their ovaries (P < 0.05). Pregnancy rates were greater in GnRH5-treated group than the GnRH5-untreated group (P < 0.01). Moreover, repeat-breeder cows bearing two CLs had a greater likelihood of pregnancy (odds ratio = 20.86) than cows bearing only one CL on their ovaries (P < 0.01). Under heat stress condition, the results highlighted that increasing luteal tissues by creating secondary CL leads to enhanced peripheral P4 concentrations and improved pregnancy outcomes in repeat-breeder dairy cows., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Egg yolk is a common cryoprotectant that can be used as a semen extender to protect the spermatozoa from damage during cryopreservation. Therefore, the aim of this study was to investigate the efficacy of fresh and lyophilized egg yolk, as a Tris-base extender, on the quality of cryopreserved goat semen. Semen from 10 rams of two different breeds (Boer and Saanen) was collected using an artificial vagina. Each ejaculate sample was divided into four equal aliquots, which contained 20% of the fresh egg yolk (a control group), and then 10%, 15%, and 20% of the lyophilized egg yolk as a Tris-base extender. Sperm motility and kinetic parameters were determined using a computer-assisted semen analyser. The results showed that the addition of 20% of the fresh egg yolk in Tris-base extender exhibited significantly higher progressive motility, progressive fast motility, distance curve line, and beat-cross frequency parameters in the post-thaw Boer and Saanen goat sperm when compared with the addition of 10%, 15%, and 20% of the lyophilized egg yolk. The percentage of total motility and immotile parameters in the post-thaw Boer and Saanen goat sperm were not significantly different between the control and 10%, 15% as well as 20% of the lyophilized egg yolk groups. Moreover, the percentage of viability parameter in the Boer and Saanen goat sperm was not significantly different between the control and 10% of the lyophilized egg yolk group but showed significant difference between the control group and 15% and 20% of the lyophilized egg yolk groups. Furthermore, the interaction between the two breeds was significantly different in terms of head activity and straightness parameter. In conclusion, the treatment with 20% of fresh egg yolk in Tris-base extender is superior to the lyophilized egg yolk. However, an addition of 10% of the lyophilized egg yolk in Tris-base extender presented the percentage of total motility and viability parameters showing no difference with 20% of fresh egg yolk. Therefore, 10% of the lyophilized egg yolk in Tris-base extender provided detail of the lyophilized egg yolk protocol in cryopreserved goat semen as an example of an alternative extender to 20% of fresh egg yolk for situations where an animal's origin represents a microbiological risk., (© 2022 Wiley-VCH GmbH.)

Sperm sexing technique is favored in the dairy industry. This research focuses on the efficiency of bovine sperm sexing using magnetic-activated cell sorting (MACS) by scFv antibody against Y-chromosome-bearing sperm (Y-scFv) coupled to magnetic microbeads and its effects on kinematic variables, sperm quality, and X/Y-sperm ratio. In this study, the optimal concentration of Y-scFv antibody coupling to the surface of magnetic microbeads was 2-4 mg/mL. PY-microbeads revealed significantly enriched Y-chromosome-bearing sperm (Y-sperm) in the eluted fraction (78.01-81.43%) and X-chromosome-bearing sperm (X-sperm) in the supernatant fraction (79.04-82.65%). The quality of frozen-thawed sexed sperm was analyzed by CASA and imaging flow cytometer, which showed that PY-microbeads did not have a negative effect on X-sperm motility, viability, or acrosome integrity. However, sexed Y-sperm had significantly decreased motility and viability. The X/Y-sperm ratio was determined using an imaging flow cytometer and real-time PCR. PY-microbeads produced sperm with up to 82.65% X-sperm in the X-enriched fraction and up to 81.43% Y-sperm in the Y-enriched fraction. Bovine sperm sexing by PY-microbeads showed high efficiency in separating Y-sperm from X-sperm and acceptable sperm quality. This initial technique is feasible for bovine sperm sexing, which increases the number of heifers in dairy herds while lowering production expenses.

This study aimed to evaluate the effects of rambutan peel powder (RP) on growth, skin mucosal and serum immunities, and immune-related gene expression of striped catfish (Pangasianodon hypophthalmus) reared in a biofloc system. Three hundred fingerlings (17.14 ± 0.12 g fish -1 ) were randomly selected and assigned to five treatments corresponding to five diets: 0 g kg -1 (control - RP0); 10 g kg -1  (RP10); 20 g kg -1  (RP20); 40 g kg -1  (RP40), and 80 g kg -1  (RP80) for 8 weeks. At weeks 4 and 8 post-feeding, growth, skin mucus, and serum immunity parameters were determined, whereas immune-related gene expressions were performed at the end of the feeding trial. Based on the results, skin mucus lysozyme (SML) and skin mucus peroxidase (SMP) were significantly higher in fish fed the RP diets compared to the control diet (P < 0.05). The highest SML and SMP levels were observed in fish fed RP40 diet, followed by RP20, RP80, RP10, and RP0. Fish-fed RP diets had higher serum lysozyme and serum peroxidase activities, with the highest value found in the RP40 diet (P < 0.05), followed by RP20, RP80, and RP10. Similarly, immune-related gene expressions (IFN2a, IFN2b, and MHCII) in the liver were significantly up-regulated in fish fed RP40. Up-regulation (P < 0.05) of IL-1, IFN2a, IFN2b, and MHCII genes was also observed in fish intestines, with the highest values observed in fish fed RP40 diet, followed by RP10, RP20, RP80, and RP0. Fish-fed diet RP diets also showed enhanced growth and FCR compared to the control, with the highest values observed in fish fed diet RP40. However, no significant differences in survival rates were found among diets. In conclusion, dietary inclusion of RP at 40 g kg -1 resulted in better growth performance, immune response, and immune related gene expressions of striped catfish (Pangasianodon hypophthalmus)., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

This study evaluates the effects of longan seed powder (LS) on the growth performance, immunological response, and immune-antioxidant related gene expression of Nile tilapia (Oreochromis niloticus). Three hundred fish (13.82 ± 0.06 g) were divided into five experiments and fed 5 diets, including the basal diet (control without LS) and basal diet containing 10 (LS10), 20 (LS20), 40 (LS40), and 80 (LS80) g kg -1 LS for eight weeks. A completely randomized design (CRD) with three replications was utilised. The growth performance and immune response were measured at weeks 4 and 8 post feeding, while the gene expressions were determined at the end of the feeding trial. The results revealed that administration of LS could significantly (P < 0.05) improve specific growth rate (SGR), weight gain (WG), and feed conversion ratio (FCR) in Nile tilapia as compared to the control group. However, no significant differences (P > 0.05) were observed in survival rates among treatments. LS-supplemented diets showed enhanced serum peroxidase activity (SPA), serum lysozyme activity (SLA), skin mucus lysozyme activity (MLA), and skin mucus peroxidase activity (MPA) at weeks 4 and 8 post-feeding, with the highest values observed in the LS20 diet (P < 0.05). Additionally, LS-supplemented diets significantly up-regulated (P < 0.05) immune and antioxidant related gene expressions (IL1, IL8, LBP, GSTa, GPX, and GSR) in the liver and intestine, with highest values observed in the LS20 treatment. The present results confirmed the beneficial effects of LS as a functional feed additive and immunostimulant for Nile Tilapia culture in a biofloc system., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Red yeast Sporidiobolus pararoseus KM281507 has been recognized as a potential feed additive. Beyond their nutritional value (carotenoids and lipids), red yeast cells (RYCs) containing high levels of β-glucan can bind mycotoxins. This study investigated the industrial feasibility of the large-scale production of RYCs, along with their ability to act as a mycotoxin binder. Under a semi-controlled pH condition in a 300 L bioreactor, 28.70-g/L biomass, 8.67-g/L lipids, and 96.10-mg/L total carotenoids were obtained, and the RYCs were found to contain 5.73% ( w / w ) β-glucan. The encapsulated RYC was in vitro tested for its mycotoxin adsorption capacity, including for aflatoxin B1 (AFB1), zearalenone (ZEA), ochratoxin A (OTA), T-2 toxin (T-2) and deoxynivalenol (DON). The RYCs had the highest binding capacity for OTA and T-2 at concentrations of 0.31-1.25 and 0.31-2.5 µg/mL, respectively. The mycotoxin adsorption capacity was further tested using a gastrointestinal poultry model. The adsorption capacities of the RYCs and a commercial mycotoxin binder (CMB) were comparable. The RYCs not only are rich in lipids and carotenoids but also play an important role in mycotoxin binding. Since the industrial-scale production and downstream processing of RYCs were successfully demonstrated, RYCs could be applied as possible feed additives.

An eight-week feeding trial was carried out to determine the effects of rambutan seed (RS) as a feed additive on the growth, skin mucus, serum immune parameters, and gene expression of Nile tilapia (Oreochromis niloticus) raised under a biofloc system. Nile tilapia fingerlings (14.77 ± 0.80 g fish -1 ) were fed five experimental diets containing 0, 5, 10, 20, and 40 g kg -1 of RS, corresponding to five treatments (RS0, RS5, RS10, RS20, and RS40) with three replications per treatment. The results showed that fish consuming the RS10 and RS20 diets presented a substantial (P < 0.05) improvement in specific growth rate (SGR), weight gain (WG), and feed conversion ratio (FCR) after eight weeks. The highest values were recorded in the RS10 diet; however, there were no significant (P > 0.05) differences exhibited in the fish survival rates between treatments. The RS supplementation diets demonstrated greater immunological parameters, particularly skin mucus and serum immune responses (P < 0.05), than that of the control after eight the eight-week feeding trial. The highest level was seen in fish fed the RS10; followed by the RS20, RS40 (P > 0.05), and RS5 diets. Regarding gene expressions, IL1, IL8, LBP, GSTa, and GSR genes were significantly up-regulated in fish provided the RS10 diet in comparison to the control and other supplemented diets (P < 0.05). However, no significant up-regulation was found in these genes among the RS0, RS5, RS20, and RS40 diets, with the exception of the GPX gene. Similarly, up-regulation of IL-8, LBP, GSTa, GPX, and GSR were noted in fish fed the RS10 diet (P < 0.05). Notably, no significant differences were evident in these genes among the RS5, RS20, and RS40 diets. In conclusion, fish fed RS10 (10 g kg -1 ) significantly enhanced growth, skin mucus, serum immunities, and immune-antioxidants related gene expressions of Nile tilapia raised under biofloc system., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

An investigation of vascularity of ovarian and uterine arteries after hormonal treatment for inactive ovaries using the short-term progesterone-based programme had not yet been explored in repeat-breeder crossbred dairy cows. To investigate the in vivo follicular and uterine arterial indices as an indicator of successful hormonal stimulation for inactive ovaries in repeat-breeder crossbred dairy cattle, 59 cows with inactive ovaries were induced with a 5-day progesterone-based protocol. At the completion of hormonal synchronisation, cows were divided into two groups according to the size of the largest follicle (LF) on their ovary: small (≤10.0 mm) and large (>10.0 mm) LFs. Vascularities of LF and uterine artery (UtA) were evaluated using a colour Doppler tool. Cows that presented with large LF had greater follicular and UtA vascular indices ( p < 0.001) and pregnancy rate ( p < 0.01) than cows bearing small LF on their ovary. There was a positive correlation ( p < 0.001) between follicular size and LF and UtA vascular indices. Our findings highlighted that in vivo LF and UtA vascular indices at the completion of hormonal stimulation might be a promising indicator for predicting success in ovarian response to hormonal stimulation for inactive ovaries of infertile crossbred dairy cows.

Nowadays, industrial poultry producers are more focused on the safety of their products, especially contaminants from feedstuffs such as mycotoxin and pesticides. The residue from animal production using antibiotic growth promoters (AGPs) may cause some problems with antimicrobial resistance in human and animals. Red yeast ( Sporidiobolus pararoseus ) has a cell wall consisting of β-glucan and mannan-oligosaccharides and pigments from carotenoids that may be suitable for use as a substitute for AGPs. The objective was to evaluate the effects of red yeast in laying hen diets on productive performance, egg quality, and duodenal health. A total of 22-week-old laying hens ( n = 480) were divided into five groups: control diet (CON), AGP at 4.5 g/kg and red yeast supplementation at 1.0 (RY1.0), 2.0 (RY2.0) and 4.0 g/kg (RY4.0) of diet. The results show that the AGP, RY2.0, and RY4.0 groups had significantly higher final body weight compared with the other groups ( p < 0.001). The red yeast supplementation improved the egg shape index ( p = 0.025), Haugh unit ( p < 0.001), and yolk color ( p = 0.037), and decreased yolk cholesterol ( p < 0.001). Diet with red yeast supplementation improved villus height to crypt depth ratio and crypt cell proliferations. In conclusion, red yeast supplementation at 2.0 g/kg of diet can substitute AGP in layer diet.

The study was executed to find out the potential effects spent coffee ground (SCG) on Nile tilapia's skin mucosal and serum immunities, disease prevention, and growth rate reared in a biofloc system. Nile tilapia fingerlings (average weight 15.25 ± 0.07 g) were disseminated into 15 aquaria (150 L tank -1 ) at a density of 20 fish per aquarium and treated five diets: SCG1 (control), SCG2 (10 g kg -1 ), SCG3 (20 g kg -1 ), SCG4 (40 g kg -1 ), and SCG5 (80 g kg -1 ) for eight weeks. A Completely Randomized Design (CRD) with three replications was applied. Growth rate, skin mucus, and serum immunities were quantified every 4 weeks; whereas the challenge study was conducted at the termination of the feeding trial. The outputs indicated that dietary incorporation of SCG give rise to the enhancement of SGR and FCR in comparison with the control, with best levels noted in fish fed SCG2 diet. Similarly, significant enhancements in skin mucosal and serum immunities were revealed in fish treated SCG2 over the control and other SCG diets. Likewise, higher survival rates against Streptococcus agalactiae were displayed in fish fed SCG, with the maximum level displayed in the fish treated SCG2. In conclusion, dietary supplementation of SCG2 (10 g kg -1 ) can be potential used as immunostimulants in tilapia aquaculture., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Fumonisin B 1 (FB 1 ) is among the most common contaminants produced by Fusarium spp. fungus from corns and animal feeds. Although FB 1 has been known to cause physical or functional defects of embryos in humans and several animal species such as Syrian hamsters, rabbits, and rodents, little is known about the precise toxicity to the embryos and the underlying mechanisms have not been fully addressed. The present study aimed to investigate its developmental toxicity and potential mechanisms of action on sphingolipid metabolism in Brown Tsaiya Ducks (BTDs) embryos. We examined the effect of various FB 1 dosages (0, 10, 20 and 40 µg/embryo) on BTD embryogenesis 72 h post-incubation. The sphingomyelin content of duck embryos decreased ( p < 0.05) in the highest FB 1 -treated group (40 µg). Failure of neural tube closure was observed in treated embryos and the expression levels of a neurulation-related gene, sonic hedgehog ( Shh ) was abnormally decreased. The sphingolipid metabolism-related genes including N -acylsphingosine amidohydrolase 1 ( ASAH1 ), and ceramide synthase 6 ( CERS6 ) expressions were altered in the treated embryos compared to those in the control embryos. Apparently, FB 1 have interfered sphingolipid metabolisms by inhibiting the functions of ceramide synthase and folate transporters. In conclusion, FB 1 -caused developmental retardation and abnormalities, such as neural tube defects in Brown Tsaiya Duck embryos, as well as are partly mediated by the disruption of sphingolipid metabolisms.

A 3-week feeding trial in a 3 × 2 × 2 factorial design was conducted with three concentrations (0, 0.5, and 5 mg/kg) of T-2 toxin (T-2) and two levels (0% and 0.5%) of green tea powder (GTP) supplements used in the diets of female brown Tsaiya ducklings (BTDs) and Kaiya ducklings (KDs), respectively. Breed had a significant effect on the growth performances and the relative weights of organs and carcass. In general, the growth performances of KDs were better than BTDs. The relative weights of organs and carcass of BTDs were typically heavier than those of KDs; however, the breast of KDs was heavier than those of BTDs. Both ducklings received 5 mg/kg of T-2 blended in the diet showed lower feed intake and body weight gain (BWG) in the second and the third week. The diet containing 5 mg/kg of T-2 and 0.5% GTP improved the BWG compared to those fed the diet supplemented with 5 mg/kg of T-2 without GTP in BTDs. Ducklings fed the diet containing 5 mg/kg of T-2 induced hypocalcemia and hypomagnesemia, as well as decreased concentrations of creatine phosphokinase and alkaline phosphatase. The concentrations of blood urea nitrogen (BUN) and glutamate oxaloacetate transaminase (GOT) were increased in KDs and BTDs fed the diet containing 5 mg/kg of T-2 without GTP, respectively. However, duckling diets containing 5 mg/kg of T-2 with 0.5% GTP lowered concentrations of BUN and GOT in the blood plasma of KDs and BTDs, respectively. The diet containing 5 mg/kg of T-2 increased the relative kidney weight but decreased the relative breast weight of ducklings. Enlarged gizzards and reduced relative leg weights were observed in BTDs fed the diets containing 5 mg/kg of T-2. In summary, BTDs are more sensitive than KDs in responding to T-2 toxicity and GTP detoxification. Green tea powder has detoxification ability and could potentially mitigate T-2 toxicity on BWG, BUN, and GOT in ducklings.

Eight weeks feeding experiment was managed to evaluate the impacts of dietary addition of pineapple peel powder (PAPP) and Lactobacillus plantarum CR1T5 (LP) individual or mixed on growth performance, skin mucus and serum immunities, as well as disease resistance of Nile tilapia. Fish (average weight 20.91 ± 0.11 g) were fed four diets: Diet 1 (0 g kg -1 PAPP and 0 CFU g -1 L. plantarum, Diet 2 (10 g kg -1 PAPP), Diet 3 (10 8  CFU g -1 L. plantarum), and Diet 4 (10 g kg -1 PAPP + 10 8  CFU g -1 L. plantarum). Serum and mucus immune responses, as well as growth rate, were assessed every 4 weeks. Ten fish were chosen for the challenge test with Streptococcus agalactiae after 8 weeks post-feeding. The findings showed that PAPP and/or LP diets increased (P ≤ 0.05) growth performance, skin mucus, and serum immune responses. The best data were obtained in fish fed a mixture of PAPP and LP. Nevertheless, no variation (P > 0.05) was recorded between groups fed PAPP or LP. The relative survival percentage (RSP, %) in Diet 2, Diet 3, and Diet 4 was 46.15%, 50.0%, and 73.08%. Fish fed mixture of PAPP + LP recorded the best (P < 0.05) survival rate versus other treatments. The current findings recommended using a mixture of PAPP and LP as promising functional additives for aquaculture practice., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

We investigated, herein, the effects of dietary inclusion of sugarcane bagasse powder (SB) on Nile tilapia development, mucosal and serum immunities, and relative immune and antioxidant genes. Fish (15.12 ± 0.04 g) were provided a basal diet (SB0) or basal diet incorporated with SB at 10 (SB10), 20 (SB20), 40 (SB40), or 80 (SB80) g kg -1 for 8 weeks. Our results demonstrated that the dietary incorporation of sugarcane bagasse powder (SB) at 20 and 40 g kg -1 significantly ameliorated FW, WG, and SGR as opposed to fish fed basal, SB10, and SB80 diets. However, no significant changes in FCR and survivability were observed between the SB supplemented diets and the control (basal diet). The mucosal immunity exhibited significantly higher SMLA and SMPA activities ( p < 0.005) in fish treated with SB diets after eight weeks. The highest SMLA and SMPA levels were recorded in fish fed SB80 followed by SB20, SB40, and SB10, respectively. For serum immunity, fish fed SB incorporated diets significantly ameliorated SL and RB levels ( p < 0.05) compared with the control. However, SP was not affected by the inclusion of SB in any diet throughout the experiment. The expression of IL1, IL8, LBP, GSTa, GPX, and GSR genes in the fish liver was significantly increased in fish fed the SB20 and SB10 diets relative to the basal diet fed fish ( p < 0.05); whereas only the IL8, LBP, and GPX genes in the intestines were substantially augmented via the SB20 and SB80 diets ( p < 0.05). IL1 and GSR were not influenced by the SB incorporated diets ( p > 0.05). In summary, sugarcane bagasse powder (SB) may be applied as a feed additive to improve growth performance, immune response, and immune and antioxidant-related gene expression in Nile tilapia.

Little is known about the degradability of mycotoxin deoxynivalenol (DON) by the spent mushroom substrate (SMS)-derived manganese peroxidase (MnP) and lignin peroxidase (LiP) and its potential. The present study investigated the growth inhibition of Fusarium graminearum KR1 and the degradation of DON by MnP and LiP extracted from SMS. The results from the 7-day treatment period showed that mycelium inhibition of F. graminearum KR1 by MnP and LiP were 23.7% and 74.7%, respectively. Deoxynivalenol production in the mycelium of F. graminearum KR1 was undetectable after treatment with 50 U/mL of MnP or LiP for 7 days. N -acetyl-D-glucosamine (GlcNAc) content and chitinase activity both increased in the hyphae of F. graminearum KR1 after treatment with MnP and LiP for 1, 3, and 6 h, respectively. At 12 h, only the LiP-treated group had higher chitinase activity and GlcNAc content than those of the control group ( p < 0.05). However, more than 60% of DON degradabilities (0.5 mg/kg, 1 h) were observed under various pH values (2.5, 4.5, and 6.5) in both MnP (50 U/g) and LiP (50 U/g) groups, while DON degradability at 1 mg/kg was 85.5% after 50 U/g of LiP treatment for 7 h in simulated pig gastrointestinal tracts. Similarly, DON degradability at 5 mg/kg was 67.1% after LiP treatment for 4.5 h in simulated poultry gastrointestinal tracts. The present study demonstrated that SMS-extracted peroxidases, particularly LiP, could effectively degrade DON and inhibit the mycelium growth of F. graminearum KR1.

A feeding trial was carried out to examine the effects of adding chestnut (Castanea sativa) polyphenols (CSP) on the growth, skin mucus and serum immune parameters of Nile tilapia (Oreochromis niloticus). Five experimental diets with inclusion levels of 0, 1, 2, 4, and 8 g kg -1 of CSP were fed to Nile tilapia fingerlings (12.77 ± 0.17 g fish -1 ) during an eight-week trial. Fish were analyzed on the fourth and eighth week to determine the influences of CSP on growth, skin mucus, and serum immune parameters. Challenging test versus Streptococcus agalactiae was evaluated at the end of the trial. Fish fed with CSP enriched diets displayed a significant increase (P ≤ 0.05) in growth and a decline in feed conversion ratio (P ≤ 0.05). Similarly, skin mucus and serum immune parameters were significantly increased (P ≤ 0.05) in fish fed CSP with respect to the control. The effects were already evident four weeks after the CSP administration. The disease protection test displayed that the fish's survival rate was significantly higher (P < 0.05) in CSP diets over the control. The relative percentage of survival (RSP) was 62.5, 75.0, 58.3, and 37.5 in fish fed diets contained 1, 2, 4, and 8 g kg -1 CSP, respectively. The best effect on growth, immune response, and disease resistance were shown in Nile tilapia fed with a diet supplementation of 2 g kg -1 CSP., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

A teratogenic agent or teratogen can disturb the development of an embryo or a fetus. Fumonisin B₁ (FB₁), produced by Fusarium verticillioides and F. proliferatum , is among the most commonly seen mycotoxins and contaminants from stale maize and other farm products. It may cause physical or functional defects in embryos or fetuses, if the pregnant animal is exposed to mycotoxin FB₁. Due to its high similarity in chemical structure with lipid sphinganine (Sa) and sphingosine (So), the primary component of sphingolipids, FB₁ plays a role in competitively inhibiting Sa and So, which are key enzymes in de novo ceramide synthase in the sphingolipid biosynthetic pathway. Therefore, it causes growth retardation and developmental abnormalities to the embryos of hamsters, rats, mice, and chickens. Moreover, maternal FB₁ toxicity can be passed onto the embryo or fetus, leading to mortality. FB₁ also disrupts folate metabolism via the high-affinity folate transporter that can then result in folate insufficiency. The deficiencies are closely linked to incidences of neural tube defects (NTDs) in mice or humans. The purpose of this review is to understand the toxicity and mechanisms of mycotoxin FB₁ on the development of embryos or fetuses.

Avian embryos are among the most convenient and the primary representatives for the study of classical embryology. It is well-known that the hatching time of duck embryos is approximately one week longer than that of chicken embryos. However, the key features associated with the slower embryonic development in ducks have not been adequately described. This study aimed to characterize the pattern and the speed of early embryogenesis in Brown Tsaiya Ducks (BTD) compared with those in Taiwan Country Chicken (TCC) by using growth parameters including embryonic crown-tail length (ECTL), primitive streak formation, somitogenesis, and other development-related parameters, during the first 72 h of incubation. Three hundred and sixty eggs from BTD and TCC, respectively, were incubated at 37.2°C, and were then dissected hourly to evaluate their developmental stages. We found that morphological changes of TCC embryos shared a major similarity with that of the Hamburger and Hamilton staging system during early chick embryogenesis. The initial primitive streak in TCC emerged between 6 and 7 h post-incubation, but its emergence was delayed until 10 to 13 h post-incubation in BTD. Similarly, the limb primordia (wing and limb buds) were observed at 51 h post-incubation in TCC embryos compared to 64 h post-incubation in BTD embryos. The allantois first appeared around 65 to 68 h in TCC embryos, but it was not observed in BTD embryos. At the 72 h post-incubation, 40 somites were clearly formed in TCC embryos while only 32 somites in BTD embryos. Overall, the BTD embryos developed approximately 16 h slower than the chicken embryo during the first 72 h of development. To our best knowledge, this is the first study to describe two distinct developmental time courses between TCC and BTD, which would facilitate future embryogenesis-related studies of the two important avian species in Taiwan.