Mubashar Hassan

Aflatoxin B1 (AFB1), a common and stable mycotoxin in feed, significantly harms poultry health and productivity by causing liver toxicity, oxidative stress, and gut microbiota imbalance. This study examined whether glucose oxidase (GO) could serve as a dietary implement to reduce AFB₁ toxicity in broilers. A total of 768 one-day-old Chinese yellow-feathered chickens were divided into four groups, with eight replicates per group and 24 chickens in each replicate, fed with a basal diet (CON), 100 µg/kg AFB₁ from contaminated culture (AFB₁), AFB₁ 100 µg/kg+300 U/kg GO (AFB₁+LGO), or AFB₁ 100 µg/kg+600 U/kg GO (AFB₁+HGO) for 64 days. The results indicated that AFB₁ exposure significantly impaired broiler growth performance, as evidenced by reduced average daily gain (ADG) and average daily feed intake (ADFI) (P<0.05). Dietary LGO supplementation effectively mitigated these adverse effects, improving both ADG and ADFI (P<0.05). Carcass characteristics, including dressing percentage, breast yield, and thigh yield, were also enhanced by LGO (P<0.05). Meat quality parameters were adversely affected by AFB₁, with increased early post-mortem pH value (45 min), drip loss, and shear force (P<0.05). LGO supplementation reversed these changes (P<0.05). AFB₁ exposure disrupted biochemical homeostasis, lowering plasma total protein and albumin levels (P<0.05) while elevating uric acid and triglyceride contents; these changes were significantly ameliorated by LGO. Additionally, AFB₁ induced oxidative stress and inflammation, as evidenced by increased nitric oxide, inducible nitric oxide synthase, and malondialdehyde levels, along with reduced antioxidant capacity (T-SOD and T-AOC) (P<0.05). GO supplementation alleviated these effects by enhancing antioxidant defenses. Furthermore, LGO modulated the gut microbiota, increasing beneficial bacterial genera (Bacteroides, Barneisiella, and Faecalibacterium) associated with improved intestinal barrier function (P<0.05). Transcriptomic and RT-qPCR analyses indicated that LGO alleviated AFB₁-induced liver injury by regulating genes (LY96, RHOB, and CCL26) linked to the TLR4 signaling pathway (P<0.05). Overall, GO exhibits hepatoprotective effects by improving antioxidant status, restoring gut-liver axis function, and modulating immune responses in broilers challenged with AFB₁.

In this study, broilers were selected as the research object to investigate the effects and mechanisms of dietary gallic acid (GA) supplementation on growth performance, meat quality, antioxidant capacity, and muscle fiber-related gene expression. A total of 750 one-day-old healthy 817 male crossbred broiler chickens were divided into five treatment groups, with six replicates per group. Birds in the control (CON) group and LPS-challenged treatment (LPS) group were fed a basal diet, and birds in the other three treatment groups received the basal diet with 150, 300, or 450 mg/kg added GA (GA150, GA300, GA450). On days 14, 17, and 20, chickens in the LPS, GA150, GA300, and GA450 groups received intramuscular injections of LPS, while chickens in the CON group received saline. The results showed that the addition of GA to the diet could effectively increase the average daily gain (ADG) of broilers from 1 to 50 days of age, and had a trend (p = 0.078) of increasing the average daily feed intake (ADFI). Adding 450 mg/kg GA to the diet significantly reduced (p < 0.05) the drip loss and pH value of pectoral muscles 45 min after slaughter, and significantly increased (p < 0.05) the lightness value of pectoral muscles 45 min post-slaughter. With an increase in GA level, the content of total volatile basic nitrogen (TVB-N) in pectoral muscles decreased linearly (p < 0.05), and the concentration of C22:6n-3 increased linearly (p < 0.05). GA effectively improved (p < 0.05) the antioxidant capacity of muscles and significantly increased (p < 0.05) the activity of total superoxide dismutase (T-SOD) in pectoral muscles after LPS stimulation, exhibiting linear and quadratic changes (p < 0.05). It also significantly increased (p < 0.05) the activity of hydrogen peroxide and decreased the activity of glutathione peroxidase (GSH-Px), while it linearly decreased (p < 0.05) the content of malondialdehyde (MDA). In addition, the dietary supplementation of GA significantly increased (p < 0.05) the expression levels of myosin heavy chain (MyHC) I and MyHC IIa in pectoral muscles and significantly decreased (p < 0.05) the expression level of MyHC IIx. In summary, the dietary addition of GA can alleviate the effect of the stress response on the growth performance of broiler chickens and improve antioxidant capacity and meat quality. The appropriate amount of dietary GA at each stage was 300 mg/kg.

In this study, broilers were selected as the research object to investigate the effects and mechanism action of dietary gallic acid (GA) supplementation on growth performance, meat quality, antioxidant capacity, and muscle fiber-related gene expression. A total of 750 one-day-old healthy 817 male crossbred broiler chickens were divided into five treatment groups, with six replicates per group: control group (CON), basal diet, basal diet with lipopolysaccharide challenge (LPS), basal diet with 150, 300, 450 mg/kg GA supplementation and LPS challenge (GA150, GA300, GA450). On days 14, 17, and 20, chickens in the LPS, GA150, GA300, and GA450 groups received intramuscular injections of LPS, while chickens in the CON group received saline. Compared with the LPS group, adding GA (GA150 and GA300) significantly increased the average daily gain (ADG) of broiler chickens from 1 to 50 days old (p < 0.05), and had the tendency to increase average daily feed intake (ADFI) (p = 0.078). Dietary addition of 450 mg/kg GA significantly reduced the pH value of breast muscle at 45min after slaughter (p < 0.05), and as the level of GA addition increased, the pH value first decreased and then increased. Compared with the LPS group, adding 450mg/kg GA significantly increased the brightness of breast muscles at 45min and 24h after slaughter (p < 0.001). Dietary supplementation of 450 mg/kg GA significantly decreased the total volatile basic nitrogen (TVB-N) content of post-mortem breast muscle (p < 0.05). Adding 450 mg/kg GA could significantly increase C22:6n-3 concentration in breast muscle (p < 0.05). Different GA levels had no significant effect on the intramuscular fat (IMF) content of broiler breast muscle (p > 0.05). Adding GA (GA150 and GA300) significantly increased (p < 0.05) the total superoxide dismutase (T-SOD) activity in the breast muscle after LPS stimulation, significantly increased (p < 0.05) the catalase (CAT) activity, and significantly reduced (p < 0.05) malonic dialdehyde (MDA) production, and had the tendency to increase glutathione peroxidase (GSH-Px) activity (p = 0.066). In addition, dietary supplementation of GA significantly increased the expression levels of myosin heavy chain (MyHC) I and MyHC IIa in breast muscles (p < 0.05), and significantly decreased the expression level of MyHC IIx (p < 0.05). In summary, dietary addition of GA can alleviate the stress response on the growth performance of broiler chickens and improve antioxidant capacity and meat quality. The appropriate amount of dietary GA at each stage was 300 mg/kg.

Deoxynivalenol (DON, Vomitoxin) is a threatening mycotoxin that mainly produces oxidative stress and leads to hepatotoxicity in poultry. Antioxidant dietary supplements dramatically boost immunity, safeguarding animals from DON poisoning. Luteolin (LUT) is an active plant-derived compound that poses influential antioxidants. This study explored the effectiveness of LUT in combination with activated charcoal (AC) in detoxifying DON in broilers. The 180 one-day broiler chickens were allocated into five different groups having six replicates in each group, provided with ad libitum feed during the trial period (28 days) as follows: in the control group, basal diet (feed with no supplementation of LUT, AC or DON); in group 2, a basal diet added with 10 mg/kg DON from contaminated culture (DON); in group 3, a basal diet augmented by 350 mg/kg LUT and DON 10 mg/kg (DON + LUT); in group 4, a basal diet supplemented by DON 10 mg/kg + AC 200 mg/kg (DON + AC); and in group 5, a basal diet supplemented by 10 mg/kg DON + 350 mg/kg LUT + 200 mg/kg AC (DON + LUT + AC). Concerning the control group, the DON-treated broilers demonstrated a significant decrease in growth performance (p < 0.05) and serum immunoglobulin (p < 0.05) contents, negatively changing the serum biochemical contents and enzymatic activities and an increase in histopathological liver lesions. Furthermore, DON substantially increased (p < 0.05) malondialdehyde (MDA) concentration and decreased total superoxide dismutase (T-SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) levels in the serum and liver. The intake of AC and LUT to the DON-contaminated diet decreased DON residue in the liver and potentially reduced the adverse effects of DON. Considering the results, supplementation of LUT with mycotoxin adsorbent has protective effects against mycotoxicosis caused by DON. It could be helpful for the development of novel treatments to combat liver diseases in poultry birds. Our findings may provide important information for applying LUT and AC in poultry production.

Aflatoxin B₁ (AFB₁) exists widely in feed and food with severe hazards, posing a serious threat to human and animal health. Epigallocatechin gallate (EGCG) and glutathione (GSH) have been reported as having anti-oxidative and other functions. The present study aimed to investigate the detoxification effect of EGCG and GSH alone or in combination on AFB₁ exposure in ducklings. Fifty one-day-old male ducklings were randomly assigned into five experimental groups (n = 10): 1. Control (CTR); 2. 0.3 mg/kg BW AFB₁ (AFB₁); 3. 0.3 mg/kg BW AFB₁ + 100 mg/kg BW EGCG (AFB₁ + EGCG); 4. 0.3 mg/kg BW AFB₁ + 30 mg/kg BW GSH (AFB₁ + GSH); 5. 0.3 mg/kg BW AFB₁ + 100 mg/kg BW EGCG + 30 mg/kg BW GSH (AFB₁ + EGCG + GSH). The experiment lasted for seven days. Compared with the CTR group, AFB₁ reduced growth performance, total serum protein and albumin content, increased serum enzyme activity (alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and γ-glutamyl transpeptidase), and caused pathological damage to the ducklings’ livers. AFB₁ exposure increased malondialdehyde content and decreased superoxide dismutase, total antioxidant capacity, catalase, glutathione peroxidase activities, and glutathione content in the liver. EGCG and GSH alone or in combination mitigated these adverse effects. Meanwhile, EGCG and GSH attenuate apoptosis of hepatocytes, and regulated AFB₁-induced changes in the abundance of genes contained in the Keap1/Nrf2 signalling and apoptotic pathways. Collectively, these results suggest that EGCG and GSH alleviate the hepatocyte injury induced by AFB₁ by inhibiting oxidative stress and attenuating excessive mitochondria-mediated apoptosis.

Background: The Smad proteins function in TGF-β signalling transduction. In the model nematodeCaenorhabditis elegans, the co-Smad, DAF-3 mediates R-Smads and performs a central role in DAF-7 signal transduction, regulating dauer formation and reproductive processes. Considering the divergent evolutionary patterns of the DAF-7 signalling pathway in parasitic nematodes, it is meaningful to explore the structure and function of DAF-3 in parasitic nematodes, such asHaemonchus contortus.Methods: Adaf-3gene (Hc-daf-3) and its predicted product (Hc-DAF-3) were identified fromH. contortusand characterised using integrated genomic and genetic approaches. In addition to immunohistochemistry employed to localiseHc-DAF-3 within adult worm sections, real-time PCR was conducted to assess the transcriptional profiles in different developmental stages ofH. contortusand RNA interference (RNAi) was performedin vitroto assess the functional importance ofHc-daf-3in the development ofH. contortus.Results: Hc-DAF-3 sequences predicted fromHc-daf-3displayed typical features of the co-Smad subfamily. The nativeHc-DAF-3 was localised to the gonad and cuticle of adult parasites. In addition,Hc-daf-3was transcribed in all developmental stages studied, with a higher level in the third-stage larvae (L3) and adult females. Moreover, silencingHc-daf-3by RNAi retarded L4 development.Conclusion: The findings of the present study demonstrated an important role ofHc-DAF-3 in the development ofH. contortuslarvae.

The complete mitochondrial (mt) genome of Trichuris skrjabini has been determined in the current study and subsequently compared with closely related species by phylogenetic analysis based on concatenated datasets of mt amino acid sequences. The whole mt genome of T. skrjabini is circular and 14,011 bp in length. It consists of a total of 37 genes including 13 protein coding genes (PCGs), two ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNAs) genes, and two non-coding regions. The gene arrangement and contents were consistent with other members of the Trichuridae family including Trichuris suis, Trichuris trichiura, Trichuris ovis, and Trichuris discolor. Phylogenetic analysis based on concatenated datasets of amino acids of the 12 PCGs predicted the distinctiveness of Trichuris skrjabini as compared to other members of the Trichuridae family. Overall, our study supports the hypothesis that T. skrjabini is a distinct species. The provision of molecular data of whole mt genome of T. skrjabini delivers novel genetic markers for future studies of diagnostics, systematics, population genetics, and molecular epidemiology of T. skrjabini.

Aflatoxin B₁ (AFB₁) is a serious threat to the poultry industry. Proanthocyanidins (PCs) demonstrates a broad range of biological, pharmacological, therapeutic, and chemoprotective properties. The aim of this study was to investigate the ameliorative effects of PCs against AFB₁-induced histopathology, oxidative stress, and apoptosis via the mitochondrial pathway in the bursa of Fabricius (BF) of broilers. One hundred forty-four one-day old Cobb chicks were randomly assigned into four treatment groups of six replicates (6 birds each replicate) for 28 days. Groups were fed on the following four diets; (1) Basal diet without addition of PCs or AFB₁ (Control); (2) basal diet supplemented with 1 mg/kg AFB₁ from contaminated corn (AFB₁); (3) basal diet supplemented with 250 mg/kg PCs (PCs); and (4) basal diet supplemented with 1 mg/kg AFB₁ + 250 mg/kg PCs (AFB₁+ PCs). The present study results showed that antioxidant enzymes activities of total superoxide dismutase (T-SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and glutathione S-transferase (GST) in AFB₁ treated group were (p < 0.05) decreased, whereas malondialdehyde (MDA) contents were significantly increased in comparison with the control group. Furthermore, we found that dietary PCs treatment ameliorated AFB₁-induced oxidative stress in the BF through inhibiting the accumulation of MDA content and enhancing the antioxidant enzymes activities (T-SOD, CAT, GSH-Px, and GST). Similarly, PCs markedly enhanced messenger RNA (mRNA) expression of antioxidant genes (SOD, CAT, GPx1, and GST) in comparison with AFB₁ group. Moreover, histological results showed that PCs alleviated AFB₁-induced apoptotic cells in the BF of broilers. In addition, both mRNA and protein expression results manifested that mitochondrial-apoptosis-associated genes (Bax, caspase-9, caspase-3, and p53 and cytochrome c) showed up-regulation, while (Bcl-2) showed down-regulation in AFB₁ fed group. The supplementation of PCs to AFB₁ diet significantly reversed the mRNA and protein expression of these apoptosis-associated genes, as compared to the AFB₁ group. Our results demonstrated that PCs ameliorated AFB₁-induced oxidative stress by modulating the antioxidant defense system and apoptosis in the BF through mitochondrial pathway in broilers.

The complete mitochondrial (mt) genome of Ostertagia trifurcata, a parasitic nematode of small ruminants, has been sequenced and its phylogenetic relationship with selected members from the superfamily Trichostrongyloidea was investigated on the basis of deduced datasets of mt amino acid sequences. The entire mt genome of Ostertagia trifurcata is circular and 14,151 bp in length. It consists of a total of 36 genes comprising 12 genes coding for proteins (PCGs), 2 genes for ribosomal RNA (rRNA), 22 transfer RNA (tRNA) genes and 2 non-coding regions, since all genes are transcribed in the same direction. The phylogenetic analysis based on the concatenated datasets of predicted amino acid sequences of the 12 protein coding genes supported monophylies of the Haemonchidae, Dictyocaulidae and Molineidae families, but rejected monophylies of the Trichostrongylidae family. The complete characterization and provision of the mtDNA sequence of Ostertagia trifurcata provides novel genetic markers for molecular epidemiological investigations, systematics, diagnostics and population genetics of Ostertagia trifurcata and its correspondents.