The efficiency of mycotoxin binding by sorbents in the in vitro model using a naturally contaminated animal feed.

Introduction: The productivity of domestic animals and the safety of food products derived from them are jeopardised by mycotoxins in animal feed. To control them, feed additives are used, which limit the absorption of mycotoxins in the gastrointestinal tract of animals by binding to them. The study aimed to evaluate the effectiveness of a new in vitro model in experiments on the binding of mycotoxins from buffers and contaminated feed and to confirm the effect of a single sorbent or mixture in binding them. Material and Methods: Nine mineral sorbents were tested for their efficiency binding eight mycotoxins. Two in vitro experiments were conducted to indicate the mycotoxin-binding capacity of sorbents, each specifying a buffer with one of two different pH levels reflecting gastrointestinal conditions (pH 3.5 and 7.0). The first investigated the sorbent with only the buffer and mycotoxin standards, while the second did so with the sorbent, buffer and feed naturally contaminated with mycotoxins (deoxynivalenol, zearalenone, and ochratoxin A). Results: The sorption was significantly lower in the trial with feed. In the first experiment at gastric pH (pH 3.5), activated charcoal bound deoxynivalenol and sepiolite bound zearalenone at 70% and 96%, respectively, whereas in the second experiment with feed, the binding was only 3% and 6%. Conclusion: The study underlines the challenge of finding a feed additive that would work comprehensively, binding all mycotoxins regulated by law.

The development of a dexamethasone challenge model for evaluating feed additives in sheep.

OBJECTIVE: To develop an accessible ruminant immune challenge model for rapid in vivo assessments of feed additives. ANIMALS: 60 hair-breed ram lambs. METHODS: Sheep were randomly assigned to 1 of 4 treatments: treatment 1, not immunosuppressed, control fed (n = 12); treatment 2, immunosuppressed, supplemented with a yeast and botanical extract (n = 18); treatment 3, immunosuppressed, supplemented with a blend of natural aluminosilicates and yeast components (n = 18); and treatment 4, immunosuppressed, control fed (n = 12). Twice-daily injections of dexamethasone (Dex; 0.1 mg/kg bodyweight, SC) were used to induce immunosuppression throughout the study (from September 25, 2020, to November 2, 2020). All sheep were immunized with keyhole limpet hemocyanin (KLH) on days 0 and 14 and injected with heat-aggregated KLH, ID, to induce a skin induration on day 15. Measurements included body weight (BW), average daily gain (ADG), CBC, and skin induration diameter. RESULTS: Dex treatment resulted in reduced BW and ADG that was not mitigated by either feed additive. Dex reduced lymphocyte percentage, RBC count, hemoglobin, hematocrit, and skin induration diameter and increased concentrations of granulocytes and granulocyte percentage. Effects on hematocrit, hemoglobin, RBC, and skin induration diameter were mitigated with the addition of feed additives. CLINICAL RELEVANCE: The described model is a tool to evaluate the ability of feed additives to mitigate the immunosuppressive effects of Dex.

Guidance on the assessment of the efficacy of feed additives.

This guidance document is intended to assist the applicant in preparing and presenting an application, as foreseen in Article 7.6 of Regulation (EC) No 1831/2003, for the authorisation of additives for use in animal nutrition. It specifically covers the assessment of the efficacy of feed additives.

Dietary supplementation of Sida rhombifolia enhances the plasma antioxidation and modulates gut microbiota in Anyi tile-like grey chickens.

Sida rhombifolia (S. rhombifolia) is a widely used herbal plant for humans because of its antioxidant and antibacterial effects, but its potential use as a feed additive for livestock has not been investigated. Twenty 350 days-old Anyi tile-like grey chickens were randomly divided into a control group (fed basal diet) and a treatment group (fed basal diet + 3% of S. rhombifolia), and these chickens were feed for 31 days. Dietary S. rhombifolia remarkably enhanced plasma antioxidants, including the significantly increased total antioxidant capability (p < 0.01), catalase (p = 0.04), and superoxide dismutase (p < 0.01) in the treatment group. Furthermore, dietary S. rhombifolia also modulated chicken cecal microbiota, including an increased microbial diversity (Shannon, p = 0.03; Chao1, p = 0.03) in the treatment group. Regarding taxonomic analysis, 34 microbial taxa showed significant differences between the two groups. Meanwhile, the dominant phylum Actinobacteriota (p = 0.04), and dominant genera Desulfovibrio (p = 0.04) and Olsenella (p = 0.02) were significantly increased after treatment, whereas the pathogenic genus Escherichia-Shigella (p = 0.04) was significantly decreased after feeding S. rhombifolia. The results indicating that S. rhombifolia has potential for use as a natural plant feed additive for chickens.

Chicken intestinal organoids: a novel method to measure the mode of action of feed additives.

There is a rapidly growing interest in how the avian intestine is affected by dietary components and feed additives. The paucity of physiologically relevant models has limited research in this field of poultry gut health and led to an over-reliance on the use of live birds for experiments. The development of complex 3D intestinal organoids or "mini-guts" has created ample opportunities for poultry research in this field. A major advantage of the floating chicken intestinal organoids is the combination of a complex cell system with an easily accessible apical-out orientation grown in a simple culture medium without an extracellular matrix. The objective was to investigate the impact of a commercial proprietary blend of organic acids and essential oils (OA+EO) on the innate immune responses and kinome of chicken intestinal organoids in a Salmonella challenge model. To mimic the in vivo prolonged exposure of the intestine to the product, the intestinal organoids were treated for 2 days with 0.5 or 0.25 mg/mL OA+EO and either uninfected or infected with Salmonella and bacterial load in the organoids was quantified at 3 hours post infection. The bacteria were also treated with OA+EO for 1 day prior to challenge of the organoids to mimic intestinal exposure. The treatment of the organoids with OA+EO resulted in a significant decrease in the bacterial load compared to untreated infected organoids. The expression of 88 innate immune genes was investigated using a high throughput qPCR array, measuring the expression of 88 innate immune genes. Salmonella invasion of the untreated intestinal organoids resulted in a significant increase in the expression of inflammatory cytokine and chemokines as well as genes involved in intracellular signaling. In contrast, when the organoids were treated with OA+EO and challenged with Salmonella, the inflammatory responses were significantly downregulated. The kinome array data suggested decreased phosphorylation elicited by the OA+EO with Salmonella in agreement with the gene expression data sets. This study demonstrates that the in vitro chicken intestinal organoids are a new tool to measure the effect of the feed additives in a bacterial challenge model by measuring innate immune and protein kinases responses.

Phytogenic feed additives as natural antibiotic alternatives in animal health and production: A review of the literature of the last decade.

The use of antibiotics in animal production raises great public safety concerns; therefore, there is an urgent need for the development of substitutes for antibiotics. In recent decades, plant-derived feed additives have been widely investigated as antibiotic alternatives for use in animal health and production because they exert multiple biological functions and are less likely to induce resistance development. This review summarizes the research history and classification of phytogenic feed additives and their main functions, potential modes of action, influencing factors, and potential negative effects. Further, we highlight the challenges in developing sustainable, safe, and affordable plant-derived antibiotic alternatives for use in livestock production.

Supplemental effects of different production methods of pine needle additives on growth performance, intestinal environment, meat quality and serum of broiler chickens.

OBJECTIVE: Pine needles are rich in many nutrients and exhibit antibacterial and antioxidant biological activities; however, the effects of different production methods of pine needle additives on the growth performance and intestinal flora of broiler chickens are not known. METHODS: Normal diets were supplemented with pine needle fermentation juice (PNF), pine needle soaking juice (PNS), or pine needle powder (PNP), and the associated effects on growth performance, relative organ weights, intestinal development, intestinal histological morphology, intestinal flora, meat quality, and serum indicators in broiler chickens were observed. RESULTS: The results showed that PNF, PNS, and PNP all significantly improved feed utilisation and promoted the growth and development of broilers. All three additives also significantly improved the structure of the intestinal flora, specifically increasing the diversity of bacteria; increasing the abundance of beneficial bacteria, such as Faecalibacterium, Rikenella, and Blautia; and decreasing the abundance of harmful bacteria, such as Staphylococcus. The antioxidant properties of pine needles were also found to intensify lipid metabolic reactions in the blood, thus leading to lower triglycerides and total cholesterol. Meanwhile, high doses of PNF reduced jejunum and ileum weights and also increased meat yellowness. Lastly, none of PNF, PNS, or PNP had an effect on relative organ weights or intestinal histological morphology. CONCLUSION: The addition of pine needles to the diet of broiler chickens can effectively promote their growth performance as well as improve their intestinal flora and serum status without side effects; in particular, the dose of 0.2% of either PNF and PNS is expected to have the capacity to replace growth-promoting antibiotics as diet additives.

Emerging microalgal feed additives for ruminant production and sustainability.

The global demand for animal-derived foods has led to a substantial expansion in ruminant production, which has raised concerns regarding methane emissions. To address these challenges, microalgal species that are nutritionally-rich and contain bioactive compounds in their biomass have been explored as attractive feed additives for ruminant livestock production. In this review, we discuss the different microalgal species used for this purpose in recent studies, and review the effects of microalgal feed supplements on ruminant growth, performance, health, and product quality, as well as their potential contributions in reducing methane emissions. We also examine the potential complexities of adopting microalgae as feed additives in the ruminant industry.

Contribution of nanotechnology to greater efficiency in animal nutrition and production.

Feed costs present a major burden in animal production for human consumption, representing a key opportunity for cost reduction and profit improvement. Nanotechnology offers potential to increase productivity by creating higher-quality and safer products. The feed sector has benefited from the use of nanosystems to improve the stability and bioavailability of feed ingredients. The development of nanotechnology products for feed must consider the challenges raised by biological barriers as well as regulatory requirements. While some nanotechnology-based products are already commercially available for animal production, the exponential growth and application of these products requires further research ensuring their safety and the establishment of comprehensive legislative frameworks and regulatory guidelines. Thus, this article provides an overview of the current state of the art regarding nanotechnology solutions applied in feed, as well as the risks and opportunities aimed to help researchers and livestock producers.

Effects of dietary supplementation of polysaccharide from Agaricus blazei Murr on productive performance, egg quality, blood metabolites, intestinal morphology and microbiota of Korean quail.

OBJECTIVE: This study aimed to investigate the effects of dietary supplementation with Agaricus blazei polysaccharide (ABP) at varying concentrations on the performance, egg quality, blood biochemistry, intestinal morphology, and microflora of quail. METHODS: The study involved a total of 2,700 Korean quails, which were randomly divided into three groups. The measured variables encompassed productive performance, egg parameters, carcass parameters, serum metabolites, immune response parameters, antioxidative properties, and gut microbiome. RESULTS: The addition of ABP did not have a significant effect on average daily feed intake. However, it was found to increase the average daily egg weight and egg production rate, reduce the feed-egg ratio. There were no significant impacts on egg quality measures such as egg shape index, egg yolk index and color, egg yolk and protein content. However, ABP supplementation significantly increased the Hough unit (p<0.01) and decreased the rate of unqualified eggs (p<0.01). Regarding serum parameters, the inclusion led to an increase in total protein concentration (p<0.05) and a reduction in low-density lipoprotein cholesterol (p<0.05). There were no significant effects observed on immune indicators such as immunoglobulin A (IgA) and IgM. ABP supplementation increased the levels of serum antioxidant indicators, including glutathione peroxidase, total superoxide dismutase (p<0.05), and total antioxidant capacity colorimeter (p<0.05). Furthermore, ABP supplementation significantly elevated the intramuscular fatty acid content in quail meat. Additionally, ABP supplementation demonstrated a significant improvement in the diversity of gut microbiota and induced alterations in the composition of the gut microbiota. CONCLUSION: The findings of this study indicate that dietary supplementation of ABP enhanced production performance and antioxidant capacity while increasing the levels of polyunsaturated fatty acids in quail muscle.

Evaluation of Increasing Dietary Concentrations of a Multi-Enzyme Complex in Feedlot Lambs' Rations.

The objective of this study was to evaluate the effects of increasing levels of the M-E complex (xylanase, glucanase, cellulase, and invertase) Optimax E® on the performance of growing lambs, their digestibility, and their rumen microbiota, and to estimate NEm, NEg, and ruminal methane levels. Forty lambs (Katahdin x Dorset; 22.91 ± 4.16 kg) were randomly assigned to dietary concentrations of ME (0, 0.2, 0.4, and 0.8% DM) and fed individually for 77 days. Increasing M-E improved feed conversion (p < 0.05) as well as NEm and NEg (p < 0.05), which were associated with increased in vivo DM and NDF digestion (linear and quadratic p < 0.01). Few microbial families showed abundancy changes (Erysipelotrichaceae, Christensenellaceae, Lentisphaerae, and Clostridial Family XIII); however, the dominant phylum Bacteroidetes was linearly reduced, while Firmicutes increased (p < 0.01), resulting in a greater Firmicutes-to-Bacteroidetes ratio. Total Entodinium showed a quadratic response (p < 0.10), increasing its abundancy as the enzyme dose was augmented. The daily emission intensity of methane (per kg of DMI or AGD) was reduced linearly (p < 0.01). In conclusion, adding the M-E complex Optimax E® to growing lambs' diets improves their productive performance by acting synergistically with the rumen microbiota, modifying the Firmicutes-to-Bacteroidetes ratio toward more efficient fermentation, and shows the potential to reduce the intensity of greenhouse gas emissions from lambs.

A Polyherbal Mixture with Nutraceutical Properties for Ruminants: A Meta-Analysis and Review of BioCholine Powder.

BioCholine Powder is a polyherbal feed additive composed of Achyrantes aspera, Trachyspermum ammi, Azadirachta indica, and Citrullus colocynthis. The objective of this study was to analyze published results that support the hypothesis that the polyherbal product BioCholine Powder has rumen bypass choline metabolites through a meta-analysis and effect size analysis (ES). Using Scopus, Web of Science, ScienceDirect, PubMed, and university dissertation databases, a systematic search was conducted for experiments published in scientific documents that evaluated the effects of BioCholine supplementation on the variables of interest. The analyzed data were extracted from twenty-one publications (fifteen scientific articles, three abstracts, and three graduate dissertations available in institutional libraries). The studies included lamb growing-finishing, lactating ewes and goats, calves, and dairy cows. The effects of BioCholine were analyzed using random effects statistical models to compare the weighted mean difference (WMD) between BioCholine-supplemented ruminants and controls (no BioCholine). Heterogeneity was explored, and three subgroup analyses were performed for doses [(4 (or 5 g/d), 8 (10 g/d)], supplementation in gestating and lactating ewes (pre- and postpartum supplementation), and blood metabolites by species and physiological state (lactating goats, calves, lambs, ewes). Supplementation with BioCholine in sheep increased the average daily lamb gain (p < 0.05), final body weight (p < 0.01), and daily milk yield (p < 0.05) without effects on intake or feed conversion. Milk yield was improved in small ruminants with BioCholine prepartum supplementation (p < 0.10). BioCholine supplementation decreased blood urea (p < 0.01) and increased levels of the liver enzymes alanine transaminase (ALT; p < 0.10) and albumin (p < 0.001). BioCholine doses over 8 g/d increased blood glucose, albumin (p < 0.10), cholesterol, total protein, and globulin (p < 0.05). The ES values of BioCholine in retained energy over the control in growing lambs were +7.15% NEm (p < 0.10) and +9.25% NEg (p < 0.10). In conclusion, adding BioCholine Powder to domestic ruminants' diets improves productive performance, blood metabolite indicators of protein metabolism, and liver health, showing its nutraceutical properties where phosphatidylcholine prevails as an alternative that can meet the choline requirements in ruminants.

Interactions between maternal parity and feed additives drive the composition of pig gut microbiomes in the post-weaning period.

BACKGROUND: Nursery pigs undergo stressors in the post-weaning period that result in production and welfare challenges. These challenges disproportionately impact the offspring of primiparous sows compared to those of multiparous counterparts. Little is known regarding potential interactions between parity and feed additives in the post-weaning period and their effects on nursery pig microbiomes. Therefore, the objective of this study was to investigate the effects of maternal parity on sow and offspring microbiomes and the influence of sow parity on pig fecal microbiome and performance in response to a prebiotic post-weaning. At weaning, piglets were allotted into three treatment groups: a standard nursery diet including pharmacological doses of Zn and Cu (Con), a group fed a commercial prebiotic only (Preb) based on an Aspergillus oryzae fermentation extract, and a group fed the same prebiotic plus Zn and Cu (Preb + ZnCu). RESULTS: Although there were no differences in vaginal microbiome composition between primiparous and multiparous sows, fecal microbiome composition was different (R2 = 0.02, P = 0.03). The fecal microbiomes of primiparous offspring displayed significantly higher bacterial diversity compared to multiparous offspring at d 0 and d 21 postweaning (P < 0.01), with differences in community composition observed at d 21 (R2 = 0.03, P = 0.04). When analyzing the effects of maternal parity within each treatment, only the Preb diet triggered significant microbiome distinctions between primiparous and multiparous offspring (d 21: R2 = 0.13, P = 0.01; d 42: R2 = 0.19, P = 0.001). Compositional differences in pig fecal microbiomes between treatments were observed only at d 21 (R2 = 0.12, P = 0.001). Pigs in the Con group gained significantly more weight throughout the nursery period when compared to those in the Preb + ZnCu group. CONCLUSIONS: Nursery pig gut microbiome composition was influenced by supplementation with an Aspergillus oryzae fermentation extract, with varying effects on performance when combined with pharmacological levels of Zn and Cu or for offspring of different maternal parity groups. These results indicate that the development of nursery pig gut microbiomes is shaped by maternal parity and potential interactions with the effects of dietary feed additives.

Natural feed additives and bioactive supplements versus chemical additives as a safe and practical approach to combat foodborne mycotoxicoses.

This review highlights the possible hazard of mycotoxins occurrence in foods and feeds in regards to foodborne diseases. The possible management of the risk of contamination of foods and feeds with mycotoxins by using natural feed additives, protecting against deleterious effects of mycotoxins or inhibiting the growth of fungi and mycotoxin production, is deeply investigated in the available literature and some effective measures for safe utilization of mycotoxin contaminated feed/food are proposed. The biological methods of decontamination, degradation or biotransformation of mycotoxins are deeply analyzed and discussed. Some natural antagonists against target fungi are also reviewed and a comparison is made with conventional fungicides for ensuring a safe prevention of mycotoxin contamination. The most common and useful chemical methods of mycotoxins decontamination of agricultural commodities or raw materials are also investigated, e.g., chemical additives inactivating or destroying and/or adsorbing mycotoxins as well as chemical additives inhibiting the growth of fungi and mycotoxin production. The practical use and safety of various kind of feed/food additives or herbal/biological supplements as possible approach for ameliorating the adverse effects of some dangerous mycotoxins is deeply investigated and some suggestions are given. Various possibilities for decreasing mycotoxins toxicity, e.g., by clarifying the mechanisms of their toxicity and using some target antidotes and vitamins as supplements to the diet, are also studied in the literature and appropriate discussions or suggestions are made in this regard. Some studies on animal diets such as low carbohydrate intake, increased protein content, calorie restriction or the importance of dietary fats are also investigated in the available literature for possible amelioration of the ailments associated with mycotoxins exposure. It could be concluded that natural feed additives and bioactive supplements would be more safe and practical approach to combat foodborne mycotoxicoses as compared to chemical additives.

Dietary Hydroxytyrosol Supplementation on Growth Performance, Gut Morphometry, and Oxidative and Inflammatory Status in LPS-Challenged Broilers.

This study assessed the effects of hydroxytyrosol (HT) on 8- to 20-day-old broilers challenged with lipopolysaccharide (LPS); 180 Cobb500™ male chicks were randomly assigned to 3 treatment groups, each comprising 10 replicates with 6 birds per replicate. Treatments included a control diet (CON), CON with LPS administration, and CON + LPS supplemented with 10 mg of HT/kg of feed. LPS was administered intraperitoneally on days 14, 16, 18, and 20. Body weight (BW), body weight gain (BWG), and the feed conversion ratio (FCR) were measured. On day 20, ten birds per treatment were slaughtered for analysis. Bursa, spleen, and liver were collected, and their respective relative weight was determined. The jejunum was destined for morphological analyses of villus height (VH), crypt depth (CD), and their ratio (VH:CD), and for mRNA expression of nuclear factor kappa B (NF-κB), catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), and interleukins 10 (IL-10), 1 beta (IL-1ß), and 8 (IL-8). HT improved BW, BWG, and FCR, and reduced crypt depth (CD) while increasing the VH:CD ratio in the jejunum. Moreover, HT downregulated mRNA expression of CAT, GPx, IL-10, and IL-1ß. In conclusion, HT enhances broiler growth performance, mitigates jejunal mucosa damage from LPS, and modulates antioxidant and immune responses.

Effect of herbal extracts in animal nutrition as feed additives.

This paper is reviewed with the objective to understand the effect of herbal extracts on animal performance as feed additives. The number of both external and internal factors which affects the production and productivity of animals obviously includes nutrition as a major factor. Feed additives are products used in animal nutrition to increase the quality of feed and animal-derived foods, as well as the performance and the health of animals. Plant extracts as feed additives are described as herbal-derived components added to ration to improve livestock performance and product quality. Many herbal extracts contain chemical components that have antioxidant, antimicrobial, anti-inflammatory, anticoccidial and anthelmintic properties to improve ruminal microbial activity, diet palatability and stimulate digestion. Bioactive chemicals found in nature, such as flavonoids and glucosinolates isoprene derivatives, are primarily responsible for the qualities of plant extracts. Plant extracts are commonly added to feed to increase palatability, productivity and to inhibit lipid oxidation. When added to meals, herbal extracts aid to decrease rancidity, delay the generation of hazardous oxidation products, and retain nutritional quality. It is concluded that; herbal extracts are important to improve growth performance and product quality.

Determination of urea in pet feed: assessing the suitability of different analytical techniques using proficiency test data.

The determination of urea in pet feed at contaminant levels using the spectrophotometric method described in Commission Regulation (EC) No 152/2009 has been reported by several EU laboratories to lack the required selectivity. Whilst urea is not authorised as an additive in pet feed, the control of urea in pet feed is of economic importance, because the addition of urea may unlawfully increase the apparent protein content. To investigate the capabilities of different analytical techniques, a proficiency test was organised where the participants (EU official control laboratories, laboratories from the academia and private laboratories) were free to use their method of choice for analysing three dog feed test materials, two samples of which were spiked with urea. Twenty-one laboratories submitted results using the following techniques: spectrophotometry (Implementing Regulation (EC) No 152/2009), LC-MS/MS, HPLC-UV, enzymatic-colorimetry, gravimetry and an 'in-house photometric' method. Only two laboratories that used LC-MS/MS were able to quantify urea accurately in the test material containing a mass fraction of 18.9 mg kg-1 whereas satisfactory results at the level of 258.9 mg kg-1 were obtained by one participant that used an 'in-house photometric method' and one that used the enzymatic method, in addition to the five participants using LC-MS/MS. The technique that provided the highest success rate across the three test materials was LC-MS/MS, whereas spectrophotometry, the enzymatic-based and HPLC-UV methods led to overestimated results in addition to a dispersion of results not suitable for compliance analysis. To address the determination of urea in pet feed at low levels, a better performing method than the one described in the legislation is required.

Knowledge Gaps on the Utilization of Fossil Shell Flour in Beef Production: A Review.

Population growth in many countries results in increased demand for livestock production and quality products. However, beef production represents a complex global sustainability challenge, including meeting the increasing demand and the need to respond to climate change and/or greenhouse gas emissions. Several feed resources and techniques have been used but have some constraints that limit their efficient utilization which include being product-specific, not universally applicable, and sometimes compromising the quality of meat. This evokes a need for novel techniques that will provide sustainable beef production and mitigate the carbon footprint of beef while not compromising beef quality. Fossil shell flour (FSF) is a natural additive with the potential to supplement traditional crops in beef cattle rations in response to this complex global challenge as it is cheap, readily available, and eco-friendly. However, it has not gained much attention from scientists, researchers, and farmers, and its use has not yet been adopted in most countries. This review seeks to identify knowledge or research gaps on the utilization of fossil shell flour in beef cattle production, with respect to climate change, carcass, and meat quality. Addressing these research gaps would be a step forward in developing sustainable and eco-friendly beef production.