CC BY
0JWPR
Journal of World's Poultry Research
2022, Scienceline Publication
J. World Poult. Res. 12(3): 133-141, September 25, 2022
Research Paper, PII: S2322455X2200015-12 License: CC BY 4.0
DOI: https://dx.doi.org/10.36380/jwpr.2022.15
Controlling Immunomodulation Effects of Deoxynivalenol Mycotoxins by NanoZinc Oxide and Probiotic in Broiler Chickens
Rasha Mahmoud Hamza Sayed-ElAhl © , Atef Abdelaziz Hassan10 , Mogda Kamel Mansour2® , Azza Moustaffa Moustaffa Abdelmoteleb © , and Ahmed Mohamed Attia El-Hamaky1 ©
1Department of Mycology and Mycotoxins, Animal Health Research Institute (AHRI), Agriculture Research Center (ARC), Giza, Egypt 2Department of Biochemistry, Toxicology and Feed Deficiency, Animal Health Research Institute (AHRI), Agricultural Research
Center (ARC), Giza, Egypt Corresponding author's E-mail: rasha_hamza2005@hotmail.com
Received: 03 July 2022 Accepted: 25 August 2022
ABSTRACT
The elimination of adverse toxic effects of mycotoxins is currently the main strategy in animal production, particularly in poultry. The current study investigated the influence of chronic administration of deoxynivalenol on the health status, biochemical and immunological parameters of broiler chickens and the efficacy of ZnO-NPs and probiotics in preventing and treating the effect of toxicity. The experiment program lasted 6 weeks and was performed on a total of 60 broiler chickens aged 5 days, divided into six groups. Group 1 received healthy feed free of toxins, group 2 was fed with deoxynivalenol (DON), group 3 received Zinc Oxide nanoparticles (ZnO-NPs) and DON, group 4 had ZnO-NPs for 1 week, then DON was added for the remaining 5 weeks, group 5 was fed on ZnO-NPs, 1 g probiotic powder/kg of diet, and DON, group 6 had ZnO-NPs and 1 g probiotic powder/kg of diet for 1 week, then DON was added for 5 weeks. The used dose of ZnO-NPs was 50 ppm, and DON was 5 ppm in the diet. The intoxicated chickens showed adverse changes as increased pro-inflammatory cytokines, serum hepatic, and pancreatic enzymes, as well as decreased free amino acids. The supplementation of ZnO-NPs and/or probiotics improved all toxic changes resulting from DON toxicity, indicating that the metal nanoparticles and probiotics can be used together in poultry feed to avoid the addition of high doses of ZnO-NPs. Therefore, the use of 50 ppm of nanomaterial supplementation plus 1 g probiotic/ kg feed for the degradation of mycotoxins in poultry feed is recommended as it is safe and affordable.
Keywords: Deoxynivalenol, Fusarium spp., Nanoparticles, Poultry, Probiotic
Worldwide morbidity and mortality from mycotoxicosis in humans and animals have a negative economic impact due to decreased health activities and output. Trichothecenes are secondary metabolites produced by Fusarium species, such as F. oxysporum, F. poae, and F. graminearum with specific references to trichothecene (T-2), deoxynivalenol (DON), and nivalenol (NIV, Chen et al., 2017; Springler et al., 2017).
Deoxynivalenol mycotoxin that has been discovered in food, may cause serious health problems and dangers to humans and animals. The toxicity occurs due to the
conjugation of epoxide group with DNA subunits, which prevents protein synthesis and causes injury to human and animal tissues (Awad et al., 2013). Acute consumption of high doses of DON may cause several gastrointestinal diseases, such as emesis and diarrhea, while chronic consumption results in growth reduction and dangerous effects on digestive organs, as well as anemia, carcinogenesis, tremors, hemorrhage, dermatitis, pulmonary edema, immunosuppression, and hormonal imbalances in humans and animals (Rotter, 1996; Pestka, 2010). Poultry production is negatively affected by DON. The mentioned results are related to the dysfunction of the gastrointestinal tract, predisposition to infectious diseases
TSQBBipaS Sayed-ElAhl RMH, Hassan AA, Mogda K Mansour, Abdelmoteleb AMM, and El-Hamaky AMA (2022). Controlling Immunomodulation Effects of Deoxynivalenol Mycotoxins by NanoZinc Oxide and Probiotic in Broiler Chickens. J. World Poult. Res., 12 (3): 133-141. DOI: https://dx.doi.org/10.36380/jwpr.2022.15
by the suppression of the cellular and humoral immune system through immune cell necrosis, serum immunoglobulins reduction, and spleen atrophy (Reddy et al., 2018; Riahi et al., 2020; Hassan et al., 2022).
Mycotoxins-management strategies have gained a lot of attention to preserve the health of animals and poultry. Although most conventional drugs are still used to treat these diseases, lower efficacy has been found due to the emergence of germ resistance, which requires finding innovative therapeutic medicines.
Nowadays, nanotechnology applications are of worldwide interest in all biomedical fields (Moghimi et al., 2005). Currently, zinc oxide nanoparticles (ZnO-NPs) have been shown to suppress microbial development, as well as enhancing appropriate antibacterial and antifungal action, and unique qualities, such as fewer environmental hazards and effectiveness in suppressing the growth of toxigenic fungi and their capacity to produce poisons (Reddy et al., 2007; Alghuthaymi et al., 2021). Moreover, immunity improvement, health promotion, and productivity in chickens can be seen as a result of ZnO-NPs (Lina et al., 2009). In addition, ZnO-NPs could couple with curcumin to diminish the viability of mycotoxin Fusarium spp. and inhibit the production of mycotoxins. A nanocomposite of ZnO-NPs and cinnamon oils may be able to remove Fusarium mycotoxins in animal feed (Gacem et al., 2021; Hassan et al., 2022).
Several studies have shown that probiotic formulations, particularly those containing Lactobacillus spp., have the potential to suppress fungal growth, and detoxify food and feed toxins (Fredua-Agyeman et al., 2017). The bio-detoxification of DON by microorganisms has been developed as a probiotic and has demonstrated significant toxin elimination benefits (Payros et al., 2016; Tiew et al., 2020; Chaudhari et al., 2022).
This study evaluated the toxicological effects of DON on broiler chickens and the efficacy of ZnO-NPs and/or probiotics in degrading the toxic effects of DON.
MATERIAL AND METHODS
Ethical approval
The experiment was approved by the Institutional Animal Health Research Ethics Committee, Egypt, and followed local laws and regulations (Vide Ref No. VMC/2014/IAEC/1046-73).
Feed samples
A total of 60 feed samples were collected from animal farms where chickens displayed toxicity
symptoms, such as vomiting, diarrhea, anorexia, weight loss, and/or abrupt death. Every 500 g of feed sample was kept in a clean plastic bag and transported to the laboratory for analysis.
Zinc nanoparticles
The ZnO-NPs had a tiny particle of 50 nm in spherical forms that were prepared at the Nuclear Research Centre, Atomic Energy Authority, Egypt.
Probiotic vials
Micro-Procell vials each included 1 g of powder containing strains of Lactobacillus plantarum (1x108 CFU), and Lactobacillus acidophilus (1x108 CFU) were provided by Cheil Bio Co., Ltd, South Korea. Saccharomyces cerevisiae (1x107 CFU) with 0.5 g skim milk carrier was provided by Lallemand, SAS, France, under the name Levucell SB 10ME Titan (LSB).
Deoxynivalenol standard solution for mycotoxin detection using Thin Layer Chromatography
Mycotoxin's standard of DON was purchased from ALDRIK Sigma Chemical Company (St. Louis, USA).
Isolation and identification of Fusarium in feed samples
Under the aseptic process, 1 g of each feed sample was placed into sterile tubes containing 9 ml sterile distilled water to obtain tenfold serial dilution. Each ml of the serial dilutions was individually put into sterile petri dish plates and mixed with Sabouraud's dextrose agar containing 0.05 mg of chloramphenicol and incubated aerobically at 25°C for 3-5 days according to ISO (2008). Fusarium species were identified by macroscopic and microscopic features of mold colonies by Pitt and Hocking (2009).
Production of deoxynivalenol mycotoxin
The cultures of mycotoxigenic Fusarium graminearum isolates were cultivated on Potato Dextrose Agar (PDA) for 5-7 days at 25°C. A total of 500 ml flasks with 100 gm of fine grounded yellow corn were autoclaved for 1 hour at 121°C. Then, slant spores of each Fusarium species of 1 week old were added to the autoclaved flask and incubated for 4 weeks at 25-28°C. The flasks were transferred to the refrigerator at 8-10°C for 2 more weeks to produce a toxin (D'Mello et al., 1998). The produced trichothecenes mycotoxins were extracted and measured by thin-layer chromatography according to
studies by Kamimura et al. (1981) and Bottalico et al. (1985).
Chickens
A total of 60 broiler chickens aged 5 days from Dyerna farm, Giza, Egypt, were kept in clean cages while healthy feed and water ad libitum were available for 6 weeks. At 7 days of age, chickens were vaccinated against Newcastle and infectious bronchitis diseases using the HIPRAVAR Hitchner B1+H120 vaccine (HIPRA, Girona, Spain) via an intra-ocular route. At 10 days of age, the vaccination against avian influenza virus was done using CEVA C® NEW FLU - KEM inactivated H5N2 vaccines (CEVA, Giza, Egypt) via subcutaneous route.
Grouping
Chickens were divided into six groups. For the first group, healthy diet was available free of toxins, the second group received only DON and kept as intoxicated chickens, the third group was fed with ZnO-NPs and DON, the fourth group received ZnO-NPs for a week, then DON was added to the ZnO-NPs, the fifth group was fed on ZnO-NPs, 1 gm probiotic powder/kg of diet, and DON, finally, the sixth group received ZnO-NPs and 1 gm probiotic powder/kg of diet for a week, then DON was added. The utilized dosage of ZnO-NPs was 50 ppm and DON was 5 ppm in the diet. The experiment lasted 6 weeks following the studies of Yalcinkya et al. (2012) and Ahmadi et al. (2014).
Sampling
At the end of the experiment, 5 ml blood samples were collected from each chicken's brachial (wing) vein using a 20-gauge needle 1 inch in length. The serum was separated by centrifugation (SHengwin, USA) for 15 minutes and immediately frozen at -20°C until analysis. According to AVMA Guidelines for the euthanasia of animals (Underwood and Anthony, 2020), Chickens were euthanized by cervical dislocation to collect livers and muscles from different groups to detect DON mycotoxin residues.
Serum biochemical parameters
Total protein was estimated according to Sonnen-Wirth and Jaret (1981). Assays for alanine aminotransferase (ALT), aspartate aminotransferase (AST), amylase, and lipase levels in serum were determined according to Lopez (2013). Malondialdehyde (MDA) and Glutathione (GSH) were determined following Ellman (1959), and Okhawa et al. (1979),
respectively. Superoxide dismutase (SOD) was estimated according to Nishikimi et al. (1972). Amino acid analysis using the Amino Acid Analyzer sykam GmbH, Analytischer Messtechnik (Gewerbering 15, D_86922 Erosing, Germany) in the Genetic Research Center, Giza,
Egypt
Cytokines detection
Interleukin 6 (IL6) and the tumor necrotic factor-a (TNF-a) were measured by Nori chicken IL6 ELISA kits and Nori chicken TNF alpha ELISA kits, respectively (Genorise scientific, Inc., Pennsylvania, USA) according to the manufacturer's protocol.
Measurement of deoxynivalenol residues
Liver and muscle tissues were subjected to direct examination using a fluorometric assay to determine Deoxynivalenol residues, according to Kongkapan et al. (2016).
Statistical analysis
The obtained data were analyzed in SPSS, version 16 using ANOVA F-test and presented as mean ± standard error. The estimation of a significant difference was set at p < 0.05 (SPSS, 2007).
RESULTS AND DISCUSSION
Probiotics improve the health and immunity of animals infected with mycotoxins through inhibiting their cellular permeability and preventing their carcinogenic effects by enhancing their biodegradation (Ghadaksaz et al., 2022).
As presented in Table 1, F. graminearum produced higher levels of DON, T-2, Diacetoxy-scirpenol (DAS), and NIV resulting in higher total trichothecenes, compared to F. sporotrichioides. These toxins produced by F. sporotrichioides and F. graminearum negatively affect immunity and inhibit protein synthesis (Black et al., 1992; Osweiler, 2000). The higher level of DON in poultry feeds was reported as 5 mg/kg according to European Commission (2006). However, the lower levels of DON sometimes resulted in adverse effects on the suppression of chicken health status (Atanasova-Penichon et al., 2012; Lucke et al., 2017). Moreover, the administration of DON mycotoxins within the range of 5-15 ppm in the chicken feed negatively influenced productive performance, metabolic markers, and spleen cell growth, leading to DNA damage (Chen et al., 2017; Riahi et al., 2020). In addition, DON mycotoxin affects lipid peroxidation and DNA damage in chicken lymphocytes (Awad et al., 2012;
Ghareeb et al., 2015). The health of the internal organs of poultry, as well as biochemical and immunological markers, are influenced by the levels of 1-5 ppm, which causes jejunal and other digestive organ anomalies in hens.
Table 2 demonstrates a decrease in antioxidant activity by DON (G2), as measured by the significant reduction of SOD activity and GSH levels compared with (G1) (p < 0.05), and an increase of oxidative stress, measured by MDA levels, compared with control group (p < 0.05). Meanwhile, the administration of ZnO-NPs and/or probiotics ameliorated the damage caused by DON as shown by the elevated SOD, GSH, and lowered MDA levels compared with (G2). Antioxidant enzymes are critical in the defense against xenobiotic-induced oxidative damage. Deoxynivalenol is a polar molecule with three free hydroxyl groups (-OH), which contributes to oxidative stress by forming a high number of free radicals with an excess of oxidation markers, such as lipid peroxidation, MDA, and depleting antioxidants enzymes,
including SOD and catalase, resulting in membrane and DNA damage (Nagy et al., 2005; Miloradovic et al., 2008; Wu et al., 2014).
Dawei et al. (2009) and Hu et al. (2012) found the ability of ZnO-NPs to scavenge free radicals and suppress the incidence of cell damage and carcinogenesis. Furthermore, the administration of ZnO-NPs enhanced the effectiveness of digestion and hence the availability of body nutrients. Moreover, Gacem et al. (2021), Mandal (2021), and Singh et al. (2021) confirmed that ZnO-NPs significantly elevated the antioxidant activity of cell membrane enzymes and reduced MDA. Probiotic strains that can restrict excessive levels of reactive radicals in vivo may prevent and treat various disorders related to oxidative stress. Probiotics elevate antioxidant defenses by producing vitamins and releasing GSH that are absorbed and dispersed throughout the body (Spyropoulos et al., 2011; Kushkevych and Jampilek, 2021; Hassan et al., 2022).
Table 1. Prevalence of trichothecenes mycotoxins produced by the isolated Fusarium spp. from poultry feeds
---^^^^Trichothecene mycotoxins (ppm) DON T-2, DAS, NIV Total trichothecene
Fusarium spp. (5 strains of each) __ (Mean ± SE) (Mean ± SE) (Mean ±SE)
F. graminearum 22.5 ± 0.60 12.5 ± 1.5 35.0 ± 2.2
F. sporotrichioides 13.5 ± 1.9 7.5 ± 0.83 21 ± 1.95
SE: Standard error, F: Fusarium, DON: Deoxynivalenol, T-2: Trichothecene, DAS: Diacetoxy-scirpenol, NIV: Nivalenol.
Table 2. Effects of dietary supplementation of ZnO-NPs and/or probiotic on oxidative stress and antioxidant activities in broiler chickens contaminated with deoxynivalenol_
Groups MDA (nmol/ml) GSH (u/ml) SOD (u/ml)
G1 1.09c ± 0.04 6.90a ± 0.06 55.52a ± 1.02
G2 1.70a ± 0.08 4.83c ± 0.13 28.00d ± 1. 89
G3 1.38b ± 0.03 5.53b ± 0.13 30.33c ± 1.45
G4 1.63a ± 0.08 5. 18bc ± 0.23 34.30c ± 1.93
G5 1.29bc ± 0.12 5.68b ± 0.18 38.33bc ± 2.73
G6 1.28bc± 0.03 5.66b ± 0.17 44.33b ± 5.99
Data are represented as mean value ± Standard error. Values in the same column with the different superscript letters are significantly different (P < 0.05). G1 : Control group, G2: Deoxynivalenol (DON) intoxicated group, G3: ZnO-NPs + DON group, G4: ZnO-NPs only for 1 week then DON + ZnO-NPs, G5: ZnO-NPs + DON + probiotic, G6: ZnO-NPs + probiotic only for 1 week then DON + ZnO-NPs + probiotic. ZnO-NPs: Zinc oxide nanoparticles, MDA: malondialdehyde, GSH: Glutathione, SOD: superoxide dismutase.
Table 3. Effects of dietary supplementation of ZnO-NPs and/or probiotic on total protein and some enzyme activities in broiler chickens contaminated with deoxynivalenol_
Groups TP (g/dl) ALT (U/l) AST (U/l) Amylase (U/l) Lipase (U/l)
G1 4.10a ± 0.07 29.00d ± 1.53 54.00d ± 0.58 881.67e ± 1.76 1.05c ± 0.03
G2 3.07c ± 0.12 54.67a ± 2.60 93.70a ± 1.82 1244.65a ± 5.53 1.25a ± 0.05
G3 3.58b ± 0.06 40.00b ± 0.58 84.33b ± 1.67 1002.67c ±29.21 1. 14bc ± 0.01
G4 3.53b ± 0.03 41.67b ± 0.33 82.63b ± 0.82 1115.67b ± 0.67 1.19ab ± 0.04
G5 3.71b ± 0.03 35.33c ± 1.20 72.37c ± 0.74 975.00cd ± 4.58 1.13bc ± 0.01
G6 3.72b ± 0.06 34.67c ± 1.45 74.67c ± 2.03 936.32d ± 18.18 1.06c ± 0.01
Data are represented as mean value ± Standard error. Values in the same column with the different superscript letters are significantly different (P < 0.05). G1 : Control group, G2: Deoxynivalenol (DON) intoxicated group, G3: ZnO-NPs + DON group, G4: ZnO-NPs only for 1 week then DON + ZnO-NPs, G5: ZnO-NPs + DON + probiotic, G6: ZnO-NPs + probiotic only for 1 week then DON + ZnO-NPs + probiotic. ZnO-NPs: Zinc oxide nanoparticles, TP: Total protein, ALT: Alanine aminotransferase, AST: Aspartate aminotransferase.
Table 3 presents the total serum protein (TP), ALT, and AST as indicators of chicken metabolism and visceral organs' health. According to Pestka, (2010), DON is a powerful protein synthesis inhibitor, causing protein turnover disruption even at low concentrations. Compared to the control group, serum ALT and AST levels have also been found to be sensitive markers of liver damage since a rise in these values in the DON treatment groups indicates leakage of injured hepatocytes following DON intake (Nyblom et al., 2004).
The pancreatic enzyme showed a significant rise in serum amylase and lipase activity after DON intake (p < 0.05). Similarly, according to previous studies, DON enhanced the activity of pancreatic chymotrypsin, amylase, and lipase (Richardson and Hamilton, 1987; Matur et al., 2010). On the contrary, Osborne et al. (1982) believe that DON ingestion causes malabsorption, defined as steatorrhea, hypocarotenoidemy, and reduces bile, pancreatic lipase, trypsin, and amylase. The administration of ZnO-NPs to the contaminated groups resulted in lower levels of liver function indicators, such as ALT and AST, compared to the non-treated group (Table 3). These findings agree with Ahmadi et al. (2014), who demonstrated that dietary ZnO-NPs decreased serum ALT in broiler chickens. ZnO-NPs also significantly reduced amylase and lipase levels (p < 0.05), resulting in a lower histopathological damage score in both the pancreas and the liver tissues.
Table 4 presents that the mycotoxicated group (G2) had a significant elevation of the pro-inflammatory cytokines, including both interleukin-6 (IL-6) and tumor necrosis factor-a (TNF-a) compared with G1 (p < 0.05). Cytokines produced by immune cells have significant functions in immune response (Giansanti et al., 2006). Similar findings to the current results were reported by Awad et al. (2013) and Hendrayani et al. (2016), who found that the administration of deoxynivalenol contaminated diet in chickens increased serum levels of IL-6 and TNF-a. Another study indicated that exposure to DON potentiated inflammatory gene expression and the chronic doses resulted in necrosis and death cells (Shi et al., 2009; Wu et al., 2015; Peng et al., 2019). Furthermore, ZnO-NPs and/or probiotics treatment in groups 3, 4, 5, and 6 showed anti-inflammatory activity by IL-6, and TNF-a. The results are consistent with earlier research by El-Bahr et al. (2020), Mandal (2021), and Sakthivel et al. (2021), who found that ZnO-NPs increased the levels of antioxidant enzymes, including SOD, catalase, and glutathione peroxidase and decreased IL-6 and TNF-a cytokines in liver and brain tissues. The current study revealed lower levels of free amino acids in G2 (Table 5). After DON exposure, the observed initial adverse effects
included reduced feed intake, emesis, diarrhea, and anorexia (Pestka and Smolinski, 2005). Moreover, DON exposure negatively affected the absorption and metabolism of different biological and intestinal flora resulting in adverse toxic effects (Awad and Zentek, 2015). It was suggested by Wu et al. (2014) that reduced levels of amino acid in contaminated chicks with DON are due to intestinal barrier dysfunction. Table 5 shows that contaminated chickens with DON (G2) adversely affected the health status of chickens by reducing levels of amino acids. However, the supplementation of ZnO-NPs and/or probiotics improved amino acids levels in groups 3, 4, 5, and 6. The amino acids are essential for protein synthesis, and reduced absorption because of DON toxicity resulted in losses in the productivity of poultry (Wu et al., 2015).
The administration of ZnO-NPs in group 3 manifested significant increase in serum amino acid of chickens, compared to the second group (p < 0.05). According to Zhang et al. (2018), leucine, isoleucine, and proline were elevated by ZnO-NPs, compared to the normal control. The serum lysine content in the ZnO-NPs and/or probiotic groups increased in comparison with the second contaminated group due to increase amino acid absorption in treated chickens (Wu et al., 2015). The present study detected that the probiotic administration in contaminated chicken feed helped the chicken overcome the toxic effects of DON on some vital organs, such as liver, this agreed with (Marzouk et al., 2008; Mehrim, 2009). Recently, it has been reported that the probiotic preparation initiated the immune function of phagocytosis and epithelial cells against DON toxicity and microbial infection (Vinderola and Ritieni, 2015; Chaudhari et al., 2022; Hassan et al., 2022). Moreover, the elimination of DON by the supplementation of probiotic strains in the feed may be due to the occurrence of malabsorption of toxins in the gastrointestinal tract of treated livestock and reduced oxidative stress and inflammatory response (Chen et al., 2013; Zhao et al., 2013; Vila-Donat et al., 2018).
Previous studies evaluated the elimination of Fusarium toxins (Trichothecenes; DON) from food using bacillus probiotics (Zhao et al., 2016; Wang et al., 2020; Thipe et al., 2022).
According to Table 6, the second group received only DON and had the highest levels of DON in liver and muscle tissues. The third group received ZnO-NPs with DON and had better elimination of DON in the liver and muscles. The trials of administration of ZnO-NPs and or probiotics in the fourth and sixth groups before the addition of DON showed lower levels of detoxification. However, the residues of DON in the liver after treatment were still within the permissible limits, as reported by Escriva et al. (2017).
Table 4. Effects of dietary supplementation of ZnO-NPs and/or probiotic IL- 6 and TNF- a: of broiler chickens contaminated with deoxynivalenol
Parameters G1 G2 G3 G4 G5 G6
IL- 6 (ng/l) 121.51d ± 2.19 147.83a ± 3.63 130.33bc ± 3.38 137.33b ± 1.67 128.33c ± 1.45 135.00bc ± 2.31
TNF- a (ng/l) 69.67d ± 2.03 112.33a ± 6.94 98.00ab ± 6.51 109.67a ± 4.84 87.67b ± 4.70 104.67ab ± 5.17
Data are represented as mean value ± Standard error. Values in the same row with the different superscript letters are significantly different (P < 0.05). G1: Control group, G2: Deoxynivalenol (DON) intoxicated group, G3: ZnO-NPs + DON group, G4: ZnO-NPs only for 1 week then DON + ZnO-NPs, G5: ZnO-NPs + DON + probiotic, G6: ZnO-NPs + probiotic only for one week then DON + ZnO-NPs + probiotic. ZnO-NPs: Zinc oxide nanoparticles. IL- 6: Interleukin-6, TNF- a: Tumor necrosis factor-a
Table 5. Effects of dietary supplementation of ZnO-NPs and/or probiotic on some serum amino acids (g/dl) of broiler chickens contaminated with deoxynivalenol
Groups Lysine Methionine Leucine Iso-leucine Arginine Proline
G1 2.09a ± 0.01 1.33a ± 0.03 2.43a ± 0.05 1.72a ± 0.03 2.62a ± 0.03 1.00a ± 0.01
G2 1.61c ± 0.05 1.11c ± 0.01 2.05b ± 0.03 1.21d ± 0.02 2.14cd ± 0.04 0.65d ± 0.04
G3 1.79b ± 0.03 1.26ab ± 0.05 2.18b ± 0.02 1.33c ± 0.00 2.21c ± 0.01 0.90b ± 0.01
G4 1.70bc ± 0.02 1.13c ± 0.02 2.09b ± 0.01 1.31c ± 0.01 2.30bc ± 0.04 0.80c ± 0.03
G5 1.74b ± 0.06 1.24b ± 0.01 2.16b ± 0.03 1.37c ± 0.03 2.38b ± 0.03 0.91b ± 0.01
G6 1.79b ± 0.03 1.22b ± 0.02 2.15b ± 0.12 1.54b ± 0.06 2.30bc ± 0.04 0.95ab ± 0.02
Data are represented as mean value ± Standard error. Values in the same column with the different superscript letters are significantly different (P < 0.05). G1: Control group, G2: Deoxynivalenol (DON) intoxicated group, G3: ZnO-NPs + DON group, G4: ZnO-NPs only for 1 week then DON + ZnO-NPs, G5: ZnO-NPs + DON + probiotic, G6: ZnO-NPs + probiotic only for one week then DON + ZnO-NPs + probiotic, ZnO-NPs: Zinc oxide nanoparticles
Table 6. Levels of deoxynivalenol residues in the liver and muscles (ppb) of contaminated broiler chickens
Organs G1 G2 G3 G4 G5 G6
Liver ND 1.56 ± 0.14 0.51 ± 0.20 0.31 ± 0.15 1.1 ± 0.27 1.3 ± 0.20
Muscles ND 1.6 ± 0.25 0.05 ± 0.01 0.03 ± 0.01 0.06 ± 0.02 0.03 ± 0.00
G1: Control group, G2: Deoxynivalenol (DON) intoxicated group, G3: ZnO-NPs + DON group, G4: ZnO-NPs only for 1 week then DON + ZnO-NPs, G5: ZnO-NPs + DON + probiotic, G6: ZnO-NPs + probiotic only for one week then DON + ZnO-NPs + probiotic DON. ZnO-NPs: Zinc oxide nanoparticles, ND: Not detectable
CONCLUSION
According to the findings, Fusarium species isolated from chicken feeds produced significant amounts of Trichothecenes mycotoxins, particularly DON. In the present investigation, this toxin has a detrimental effect on the health status, immunological state, and output of chickens. Furthermore, supplementation of ZnO-NPs, either alone or in combination with the probiotic, showed considerable potential in eliminating the negative activity of DON in broiler chickens. As a result, supplementation of ZnO-NPs and probiotic can significantly improve poultry and animal health, as well as their products such as meat, eggs, milk, wool, and leather. Finally, it is recommended to conduct future studies on the addition of probiotics to ZnO-NPs for the degradation of mycotoxins
and lowering the used doses of nanomaterials to avoid their toxic levels.
DECLARATIONS
Acknowledgments
Authors would like to thank Prof. Dr. H. H. Mansour, Head of Central Laboratory of Elemental and Isotopic Analysis, Nuclear Research Centre, Atomic Energy Authority, Egypt, for helping us in the preparation and characterization of zinc nanoparticles.
Authors' contribution
Rasha Sayed-ElAhl designed the research. Rasha Sayed-ElAhl, Atef Hassan, Mogda Mansour, Azza Abdelmoteleb, and Ahmed El Hamaky performed the experimental duties of this study and analyzed the data. Mogda Mansour did the statistical analyses. Atef Hassan and Mogda Mansour wrote the draft of the manuscript.
Mogda Mansour, Rasha Sayed-ElAhl, Azza Abdelmoteleb, and Ahmed El Hamaky have taken part in the revision of the manuscript. All authors read and approved the final version of the manuscript.
Competing interests
The authors declare that they have no competing interests.
Ethical considerations
The authors investigated ethical issues such as plagiarism, permission to publish, malfeasance, data falsification and/or fabrication, double publishing and/or submission, and redundancies.
REFERENCES
Ahmadi F, Ebrahimnezjad Y, Ghalehkandi JG, and Sis NM (2014). The effect of dietary zinc oxide nanoparticles on the antioxidant state and serum enzymes activity in broiler chickens during starter stage. In: International Conference on Biological, Civil and Environmental Engineering. Dubai (UAE): pp. 26-28. DOI: https://www.doi.org/10.15242/iicbe.c0314120
Alghuthaymi MA, Hassan AA, Kalia A, Sayed El Ahl RMH, El Hamaky AAM, Oleksak P, Kuca K, and Abd-Elsalam KA (2021). Antifungal nano-therapy in veterinary medicine: Current status and future prospects. Journal of Fungi, 7(7): 494. DOI: https://www.doi .org/10.3390/j of7070494
Atanasova-Penichon V, Pons S, Pinson-Gadais L, Picot A, Marchegay G, Bonnin-Verdal MN, Ducos C, Barreau C, Roucolle J, Sehabiague P et al. (2012). Chlorogenic acid and maize ear rot resistance. A dynamic study investigating Fusarium graminearum development, deoxynivalenol production, and phenolic acid accumulation. Molecular Plant-Microbe Interactions, 25(12): 1605-1616. DOI: https://www.doi.org/10.1094/MPMI-06-12-0153-R
Awad WA, Ghareeb K, Dadak A, Gille L, Staniek K, Hess M, and Böhm J (2012). Genotoxic effects of deoxynivalenol in broiler chickens fed with low protein diets. Poultry Science, 91(3): 550-555. DOI: https://www.doi .org/10.3382/ps.2011-01742
Awad WA and Zentek J (2015). The feed contaminant deoxynivalenol affects the intestinal barrier permeability through inhibition of protein synthesis. Archives of Toxicology, 89(6): 961-965. DOI: https://www.doi .org/10.1007/s00204-014-1284-9
Awad W, Ghareeb K, Bohm J, and Zentek J (2013). The toxicological impacts of the Fusarium mycotoxin, deoxynivalenol, in poultry flocks with special reference to immunotoxicity. Toxins, 5(5): 912925. DOI: https://www.doi.org/10.3390/toxins5050912
Black RD, McVey DS, and Oeheme FW (1992). Inmunotoxcicity in the bovine animal: A review. Veterinary and Human Toxicology Journal, 34(5): 438-442. Available at:
https://europepmc.org/ article/med/1455614
Bottalico A, Visconti A, Logrieco A, Sol-frizzo M, and Mirocha CJ (1985). Occurrence of zearalenols (diasteromeric mixture) in corn stalk rot and their production in associated Fusarium species. Applied and Environmental Microbiology Journal, 49(3): 547-551. DOI: https://doi.org/10.1128%2Faem.49.3.547-551.1985
Chaudhari A, Bharti A, and Dwivedi MK (2022). Role of probiotics in the management of fungal infections. Probiotics in the Prevention and Management of Human Diseases, Chapter 21, pp. 305-330. DOI: https://www.doi.org/10.1016/B978-0-12-823733-5.00015-5
Chen SS, Li YH, and Lin MF (2017). Chronic exposure to the Fusarium mycotoxin deoxynivalenol: Impact on performance, immune organ,
and intestinal integrity of slow-growing chickens. Toxins, 9(10): 334. DOI: https://www.doi.org/10.3390/toxins9100334
Chen L, Liu W, Li Y, Luo S, Liu Q, Zhong Y, Jian Z, and Bao M (2013). Lactobacillus acidophillus ATCC 4356 attenuates the atherosclerotic progression through modulation of oxidative stress and inflammatory process. International
Immunopharmacology, 17(1): 108-115. DOI:
https://www.doi.org/10.1016/j.intimp.2013.05.018
Dawei A, Zhisheng W, and Anguo Z (2009). Protective effects of Nano-ZnO on the primary culture mice intestinal epithelial cells in in vitro against oxidative injury. Journal of Animal and Veterinary Advances, 8(10): 1964-1967. Available at:
https://medwelljournals.com/abstract/?doi=javaa.2009.1964.1967
D'Mello JPF, Macdonald AMC, Postel D, Dijksma WTP, Dujardin A, and Placinta CM (1998). Pesticide use and mycotoxin production in Fusarium and phytopathogens. European Journal of Plant Pathology, 104: 741-751. 1DOI:
https://www.doi.org/10.1023/A:1008621505708
El-Bahr SM, Shousha S, Albokhadaim I, Shehab A, Khattab W, Ahmed-Farid O, El-Garhy O, Abdelgawad A, El-Naggar M, Moustafa M et al. (2020). Impact of dietary zinc oxide nanoparticles on selected serum biomarkers, lipid peroxidation and tissue gene expression of antioxidant enzymes and cytokines in Japanese quail. BMC Veterinary Research, 16(1): 394. DOI:
https://www.doi.org/10.1186/s12917-020-02482-5
Ellman GL (1959). Tissue sulphydryl groups. Archives of Biochemistry and Biophysics, 82(1): 70-77. DOI: https://www.doi.org/10.1016/0003-9861(59)90090-6
Escriva L, Font G, Manyes L, and Berrada H (2017). Studies on the presence of mycotoxins in biological samples: An overview. Toxins, 9(8): 251. DOI:
http://www.doi:10.3390/toxins9080251
European Commission (2006). The presence of deoxynivalenol, zearalenone, ochratoxin A, T-2, and HT-2 and fumonisins in products intended for animal feeding. Official Journal of the European Union, 299: 7-9. Available at: http://data.europa.eu/eli/reco/2006/576/oj
Fredua-Agyeman M, Stapleton P, Basit AW, Beezer AE, and Gaisford S (2017). In vitro inhibition of Clostridium difficile by commercial probiotics: A microcalorimetric study. International Journal of Pharmaceutics, 517(1): 96-103.
DOI: https://www.doi.org/10.1016/j.ijpharm.2016.12.005
Gacem MA, Terzi V, and Ould-El-Hadj-Khelil A (2021). Zinc nanostructures: Detection and elimination of toxigenic fungi and mycotoxins. Zinc-based nanostructures for environmental and agricultural applications, Chapter 20, pp. 403-430. DOI: https://www.doi.org/10.1016/B978-0-12-822836-4.00006-9
Ghadaksaz A, Nodoushan SM, Sedighian H, Behzadi E, and Fooladi AAI (2022). Evaluation of the role of probiotics as a new strategy to eliminate microbial toxins: A review. Probiotics and Antimicrobial Proteins, 14: 224-237. DOI: https://www.doi.org/10.1007/s12602-021-09893-2
Ghareeb K, Awad WA, Zebeli Q, and Böhm J (2015). Deoxynivalenol in chicken feed alters the vaccinal immune response and clinical biochemical serum parameters but not the intestinal and carcass characteristics. Journal of Animal Physiology and Animal Nutrition, 100(1): 53-60.
DOI: https://www.doi.org/10.1111/jpn.12328
Giansanti F, Giardi MF, and Botti D (2006). Avian cytokines - an overview. Current Pharmaceutical Design, 12(24): 3083-3099. DOI: https://www.doi.org/10.2174/138161206777947542
Hassan AA, Sayed-ElAhl RMH, ElHamaky AM, Oraby NH, and Barakat MH (2022). Probiotics in the prevention and control of foodborne diseases in humans. probiotics in the prevention and management of human diseases, Chapter 25, pp. 363-382. DOI: https://www.doi.org/10.1016/C2020-0-00028-9
Hendrayani SF, Al-Harbi B, Al-Ansari MM, Silva G, and Aboussekhra A (2016). The inflammatory/cancer-related IL-6/STAT3/NF-kB positive feedback loop includes AUF1 and maintains the active state of breast myofibroblasts. Oncotarget, 7(27): 41974-41985. DOI: https://www.doi.org/10.18632/oncotarget.9633
Hu CH, Gu LY, Luan ZS, Song J, and Zhu K (2012). Effects of montmorillonite-zinc oxide hybrid on performance, diarrhea, intestinal permeability, and morphology of weanling pigs. Animal Feed Science and Technology, 177(1-2): 108-115. DOI: https://www.doi.org/10.1016/j.anifeedsci.2012.07.028
International Standards Organization (ISO) (2008). Microbiology of food and animal feeding stuffs - horizontal method for the enumeration of yeasts and moulds - Part 1: Colony count technique in products with water activity greater than 0, 95. International Organization for Standardization, ISO 21527-1: 2008. Available at: https://www.iso.org/standard/38275.html
Kamimura H, Nishijima M, Yasuda K, Saito K, Ibe A, Nagayama T, Ushiyama H, and Naoi Y (1981). Simultaneous detection of several Fusarium mycotoxins in cereals, grains, and foodstuffs. Journal of the Association of Official Analytical Chemists, 64(5): 1067-1073. Available at: https://pubmed.ncbi.nlm.nih.gov/7287603/
Kongkapan J, Giorgi M, Poapolathep S, Isariyodom S, and Poapolathep A (2016). Toxicokinetics and tissuedistribution of nivalenol in broiler chickens. Toxicon, 111: 31-36. DOI: https://www. doi.org/10.1016/j .toxicon.2015.12.013
Kushkevych I and Jampilek J (2021). Effect of intestinal microbiome, antibiotics, and probiotics in the prevention and management of ulcerative colitis. Probiotics in the prevention and management of human diseases. Academic Press. Chapter 4, pp. 59-92. DOI: https://www.doi.org/10.1016/B978-0-12-823733-5.00016-7
Lina T, Jianyang J, Fenghua Z, Huiying R and Wenli L (2009). Effect of nano-zinc oxide on the production and dressing performance of broiler. Chinese Agricultural Science Bulletin, 2(3): 318. Available at: http://www.casb.org.cn/EN/abstract/abstract5231 .shtml
Lopez J (2013). Carl A. Burtis, Edward R. Ashwood and David E. Bruns (eds): Tietz Textbook of Clinical Chemistry and Molecular Diagnosis, 5th Edition. Indian Journal of Clinical Biochemistry, 28: 104-105. DOI: https://www.doi.org/10.1007/s12291-012-0287-7
Lucke A, Doupovec B, Paulsen P, Zebeli Q, and Böhm J (2017). Effects of low to moderate levels of deoxynivalenol on feed and water intake, weight gain, and slaughtering traits of broiler chickens. Mycotoxin Research, 33(4): 261-271. Avaialble at: https://www.doi.org/10.1007/s12550-017-0284-z
Mandal TK (2021). ZnO nanostructures for food packaging applications. Zinc-based nanostructures for environmental and agricultural applications. Chapter 1, pp. 367-384. DOI: https://www.doi.org/10.1016/B978-0-12-822836-4.00024-0
Marzouk MS, Moustafa MM, and Mohamed NM (2008). The influence of some probiotics on the growth performance and intestinal microbial flora of Oreochromisniloticus. Proceedings of 8th International Symposium on Tilapia in Aquaculture, Cairo, Egypt, pp. 1059-1071. Available at:
https://cals.arizona.edu/azaqua/ista/ISTA8/Mrzouk.pdf
Matur E, Ergul E, Akyazi I, Eraslan E, and Cirakli ZT (2010). The effects of Saccharomyces cerevisiae extract on the weight of some organs, liver, and pancreatic digestive enzyme activity in breeder hens fed diets contaminated with aflatoxins. Poultry Science, 89(10): 22132220. DOI: https://www.doi .org/10.3382/ps.2010-00821
Mehrim AI (2009). Effect of dietary supplementation of Biogen (Commercial probiotic) on mono-sex Nile tilapia Oreochromisniloticus under different stocking densities. Journal of Fisheries and Aquatic Sciences, 4(6): 261-273. DOI: https://www.doi.org/10.3923/jfas.2009.261.273
Miloradovic Z, Jovovic D, Mihailovic-Stanojevic N, Milanovic JG, and Milanovic S (2008). Effects of long-term losartan and L-arginine treatment on haemodynamics, glomerular filtration, and sod activity
in spontaneously hypertensive rats. Canadian Journal of Physiology and Pharmacology, 86(4): 210-214. DOI:
https://www.doi.org/10.1139/Y08-022
Moghimi SM, Hunter AC, and Murray JC (2005). Nanomedicine: Current status and future prospects. The FASEB Journal, 19(3): 311-330. DOI: https://www.doi .org/10.1096/fj .04-2747rev
Nagy CM, Fejer SN, Berek L, Molnar J, and Viskolcz B (2005). Hydrogen bondings in deoxynivalenol (DON) conformations a density functional study. Journal of Molecular Structure: THEOCHEM, 726(1-3): 55-59. DOI:
https://www.doi.org/10.1016/j.theochem.2005.02.079
Nishikimi M, Rao NA, and Yagi K (1972). The occurrence of superoxide anion in the reaction of reduced phenazine methosulfate and molecular oxygen. Biochemical and Biophysical Research Communications, 46(2), 849-854. DOI:
https://doi.org/10.1016/S0006-291X(72)80218-3
Nyblom H, Berggren U, Balldin J, and Olsson R (2004). High AST/ALT ratio may indicate advanced alcoholic liver disease rather than heavy drinking. Alcohol and Alcoholism, 39(4): 336-339. DOI: https://www.doi.org/10.1093/alcalc/agh074
Okhawa H, Ohisi N, and Yagi K (1979). Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry, 95: 351-358. DOI:
https://www.doi.org/10.1016/0003-2697(79)90738-3
Osborne DJ, Huff WE, Hamilton PB, and Burmeister HR (1982). Comparison of ochratoxin, aflatoxin, and T-2 toxin for their effects on selected parameters related to digestion and evidence for specific metabolism of carotenoids in chickens. Poultry Science, 61(8): 1646-1652. DOI: https://www.doi.org/10.3382/ps.0611646
Osweiler GD (2000). Mycotoxins-contemporary issues of food animal health and productivity. Veterinary Clinics of North America: Food Animal Practice, 16(3): 511-530. DOI:
https://www.doi.org/10.1016/S0749-0720(15)30084-0
Payros D, Alassane-Kpembi I, Pierron A, Loiseau N, Pinton P, and Oswald IP (2016). Toxicology of deoxynivalenol and its acetylated and modified forms. Archives of Toxicology, 90: 2931-2957. DOI: https://www.doi.org/10.1007/s00204-016-1826-4
Peng Z, Liao Y, Chen L, Liu S, Shan Z,_Nüssler AK, Yao P, Yan H, Liu L, and Yang W (2019). Heme oxygenase-1 attenuates low-dose of deoxynivalenol-induced_hver inflammation potentially associating with microbiota. Toxicology and Applied Pharmacology, 374: 2031. DOI: https://www.doi.org/10.1016/j.taap.2019.04.020
Pestka J (2010). Toxicological mechanisms and potential health effects of deoxynivalenol and nivalenol. World Mycotoxin Journal, 3(4): 323347. DOI: https://www.doi.org/10.3920/WMJ2010.1247
Pestka JJ and Smolinski AT (2005). Deoxynivalenol: Toxicology and potential effects on humans. Journal of Toxicology and Environmental Health, Part B, 8(1): 39-69. DOI: https://www.doi.org/10.1080/10937400590889458
Pitt JI and Hocking AD (2009). Fungi and food spoilage. Third Edition. Published by Springer Dordrech Heidelberg, London, New York, 519: 19-52. DOI: https://www.doi.org/10.1007/978-0-387-92207-2
Reddy KE, Song J, Lee HJ, Kim M, Kim DW, Jung HJ, Kim B, Lee Y, Yu D, Kim DW et al. (2018). Effects of high levels of deoxynivalenol and zearalenone on growth performance, and hematological and immunological parameters in pigs. Toxins, 10(3): 114. DOI: https://www.doi.org/10.3390/toxins10030114
Reddy KM, Feris K, Bell J, Wingett DG, and Hanley C (2007). Selective toxicity of zinc oxide nanoparticles to prokaryotic and eukaryotic systems. Applied Physics Letters, 90(21): 2139021. DOI: https://www.doi.org/10.1063/L2742324
Riahi I, Marquis V, Ramos AJ, Brufau J, Esteve-Garcia E, and Pérez-Vendrell AM (2020). Effects of deoxynivalenol-contaminated diets on productive, morphological, and physiological indicators in
broiler chickens. Animals, 10(10): 1795. DOI: https://www.doi.org/10.3390/ani10101795
Richardson KE and Hamilton PB (1987). Enhanced production of pancreatic digestive enzymes during aflatoxicosis in egg-type chickens. Poultry Science, 66(4): 640-644. DOI: https://www.doi .org/10.3382/ps.0660640
Rotter BA (1996). Invited review: Toxicology of deoxynivalenol (vomitoxin). Journal of Toxicology and Environmental Health, 48(1): 1-34. DOI: https://doi.org/10.1080/009841096161447
Sakthivel S, Periakaruppan R, Chandrasekaran R, and Abd-Elsalam KA (2021). Zinc nanomaterials: Synthesis, antifungal activity, and mechanisms. Zinc-Based Nanostructures for Environmental and Agricultural Applications. Chapter 8, pp. 139-165. DOI: https://doi.org/10.1016/B978-0-12-822836-4.00009-4
Shi Y, Porter K, Parameswaran N, Bae HK, and Pestka JJ (2009). Role of GRP78/BiP degradation and ER stress in deoxynivalenol-induced interleukin-6 upregulation in the macrophage. Toxicological Sciences, 109(2): 247-255. DOI:
https://www.doi .org/10.1093/toxsci/kfp060
Singh NB, Agarwal A, and Uzo-God OC (2021). Zinc nanomaterials: A safe animal feed supplement. Zinc-Based Nanostructures for Environmental and Agricultural Applications. Chapter 19, pp. 385401. DOI: https://www.doi.org/10.1016/b978-0-12-822836-4.00026-4
Sonnen-Wirth AC and Jaret L (1981). Gradwal's clinical laboratory methods and diagnosis. Immunology, 43(4): 803. Available at: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1555089/
Springler A, Hessenberger S, Reisinger N, Kern C, Nagl V, Schatzmayr G, and Mayer E (2017). Deoxynivalenol and its metabolite deepoxy-deoxynivalenol: multi-parameter analysis for the evaluation of cytotoxicity and cellular effects. Mycotoxin Research, 33: 25-37. DOI: https://www.doi.org/10.1007/s12550-016-0260-z
Statistical Package for the Social Sciences (SPSS) (2007). Social Scientist Statistical Package for Windows Release 16.0. SPSS Inc.
Spyropoulos BG, Misiakos EP, Fotiadis C, and Stoidis CN (2011). Antioxidant properties of probiotics and their protective effects in the pathogenesis of radiation-induced enteritis and colitis. Digestive Diseases and Sciences, 56(2): 285-294. DOI: https://www.doi.org/10.1007/s10620-010-1307-1
Thipe VC, Mentor S, Lima CS, Freitas LF, Fonseca AC, Nogueira KM, Rodrigues AS, Batista JG, Ferreira AH, and Lugäo AB (2022). The role of probiotics in maintaining immune homeostasis. In Probiotics in the Prevention and Management of Human Diseases. Academic Press. Chapter 3, pp. 41-58. DOI: https://www.doi.org/10.1016/B978-0-12-823733-5.00024-6
Tiew PW, Mac Aogain MM, Binte Mohamed NAT, Thng KX, Goh K, Lau KJ, and Chotirmall SH (2020). The mycobiome in health and disease: Emerging concepts, methodologies, and challenges.
Mycopathologia, 185(2): 207-231. DOI:
https://www.doi.org/10.1007/s11046-019-00413-z
Underwood W and Anthony R (2020). AVMA guidelines for the euthanasia of animals: 2020 Edition. Retrieved on March 2013, 30: pp. 2020-1. Available at:
https://www.avma.org/sites/default/files/2020-02/
Vila-Donat P, Marin S, Sachis V, and Ramos AJ (2018). A review of the mycotoxin adsorbing agents, with an emphasis on their multi-binding capacity, for animal feed decontamination. Food and chemical toxicology, 114: 246-259. DOI: https://www.doi.org/10.1016/j.fct.2018.02.044
Vinderola G and Ritieni A (2015). Role of probiotics against mycotoxins and their deleterious effects. Journal of Food Research, 4(1): 10-21. DOI: https://www.doi.org/10.5539/jfr.v4n1p10
Wang S, Hou Q, Guo Q, Zhang J, Sun Y, Wei H, and Shen L (2020). Isolation and characterization of a deoxynivalenol-degrading bacterium Bacillus licheniformis YB9 with the capability of modulating intestinal microbial flora of mice. Toxins, 12(3): 184. DOI: _ https://www.doi.org/10.3390/toxins12030184
Wu L, Liao P, He L, Ren W, Yin J, Duan J, and Li T (2015). Growth performance, serum biochemical profile, jejunal morphology, and the expression of nutrients transporter genes in deoxynivalenol (DON)-challenged growing pigs. BMC Veterinary Research, 11(1): 144. DOI: https://www.doi .org/10.1186/s12917-015-0449-y
Wu QH, Wang X, Yang W, Nussler AK, Xiong LY, Kuca K, Dohnal V, Zhang XJ, and Yuan ZH (2014). Oxidative stress-mediated cytotoxicity and metabolism of T-2 toxin and deoxynivalenol in animals and humans: An update. Archives of Toxicology, 88(7): 1309-1326. DOI: https://www.doi .org/10.1007/s00204-014-1280-0
Yalcinkya L, Cinar M, Yildrim E, Erat S, Basalan M, and Gungor T (2012). The effect of probiotic and organic zinc alone and in combination in broiler diets on the performance and some blood parameters. Italian Journal Animal Science, 11(3): 297-302. DOI: https://www.doi.org/10.4081/ijas.2012.e55
Zhang W, Zhao Y, Li F, Li L, Feng Y, Min L, Ma D, Yu S, Liu J, Zhang H et al. (2018). Zinc oxide nanoparticle caused plasma metabolomic perturbations correlate with hepatic steatosis. Frontiers in Pharmacology, 9: 57. DOI: https://www.doi.org/10.3389/fphar.2018.00057
Zhao L, Li X, Ji C, Rong X, Liu S, Zhang J, and Ma Q (2016). Protective effect of Devosia sp. ANSB714 on growth performance, serum chemistry, immunity function and residues in kidneys of mice exposed to deoxynivalenol. Food and Chemical Toxicology, 92: 143-149. DOI: https://www.doi.org/10.1016/j.fct.2016.03.020
Zhao Y, Zhao L, Zheng X, Fu T, Guo H, and Ren F (2013). Lactobacillus salivarius strain FD 889 induced longevity in caenorhabditiselegans by dietary restriction. Journal of Microbiology, 51(2): 183-188. DOI: https://www.doi.org/10.1007/s12275-013-2076-2
