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Calibrin®-A Safeguards Ducklings and Broiler Chicks Against Aflatoxin

Young animals (e.g., ducklings and broiler chicks) are particularly susceptible to the harmful and production-limiting effects of aflatoxins. Therefore, it’s important for producers to have preventative tactics in place to reduce the risk of aflatoxicosis in their flock. Peer-reviewed studies have shown that an all-natural mineral-based feed additive, Calibrin®-A (available in select international markets), can help ameliorate the effects of aflatoxin on ducklings and broiler chicks when fed from hatch.

Biochemical and Biological Aflatoxin Effects

Aflatoxins are produced by the fungi Aspergillus flavus and A. parasiticus. Multiple metabolite forms are produced by Aspergillus, including B1, B2, G1 and G2, with aflatoxin B1 (AFB1) the most common and potent cause of aflatoxicosis. In the right environment, typically hot and humid conditions, aflatoxins can be produced in Aspergillus-contaminated feed.

Subclinical cases of aflatoxicosis generally cause reduced weight gain and productivity, while more severe cases can cause liver damage, immunosuppressive effects, gastrointestinal dysfunction and mortality. Various biological and biochemical parameters can be measured to determine the effects aflatoxin has on the body, including growth performance, intestinal morphology, serum biochemistry and oxidative stress.

A Natural Aflatoxin Binder

These same biological and biochemical parameters can also be used to assess the efficacy of feed additives, like all-natural Calibrin-A, in protecting animals against the harmful and profit-limiting effect of aflatoxins. Calibrin-A rapidly adsorbs polar mycotoxins like aflatoxin, ergotamine and ergovaline, due to a combination of its natural properties and Amlan’s proprietary thermal processing technique. By binding aflatoxin in the gut, Calibrin-A prevents aflatoxin from being absorbed into the bloodstream and exerting its toxic effects.

The effectiveness of Calibrin-A in protecting young birds, and therefore producers’ profits, has been reported in a number of peer-reviewed journal articles and scientific conference presentations, three of which are described below.

Calibrin-A Alleviates Aflatoxin Effects in Ducklings

Two papers, one published in the Journal of Applied Poultry Research and the other in Poultry Science, reported the biological and biochemical effects of aflatoxin (AFB1, from naturally contaminated corn) on ducklings from 1 to 21 days of age. The first report looked at four diets (with and without aflatoxin, with and without Calibrin-A) and the effects of these diets on hematology and serum biochemistry.1 To look at the effects of aflatoxin on growth performance and liver and intestinal health, the second report used the results from the first study, as well as four additional treatments that used two lower concentrations of aflatoxin (eight treatments total, Table 1). 2

Table 1: Aflatoxin concentration of “clean” and naturally contaminated corn-based diets fed to ducklings with or without Calibrin-A.

Two control diets containing “clean” corn, with and without 0.1% Calibrin-A, were used in the experiment to determine if Calibrin-A had any negative effects on growth performance. The control diets did have detectable levels of aflatoxin; however, they were considered “clean” because these concentrations were below the tolerable level of aflatoxin contamination (20 ug/kg) set by some regulators. Despite careful ingredient selection, the control diets did have aflatoxin contamination, which demonstrates the importance of having a risk-management strategy in place to prevent mycotoxin-related health and performance issues in your flock.

Feeding 0.1% Calibrin-A in the clean diet did not change the feed conversion ratio with values of 1.59 and 1.57 for 0 and 0.1% Calibrin-A, respectively.1 This indicates that adding Calibrin-A to the ration did not interfere with the digestibility and utilization of nutrients needed for normal growth.1

The high aflatoxin diet had lower average daily gain (ADG) compared to the Calibrin-A clean diet and greater mortality compared to negative control clean diet.2 Increasing the amount of contaminated corn had a linear, quadratic or both effect on reducing ADG and increasing mortality. However, Calibrin-A decreased mortality irrespective of the contaminated-corn concentration, and the variability in growth that significantly increased at the 50 and 100% aflatoxin level in the second report was also alleviated by the addition of Calibrin-A.2

As expected, aflatoxin caused intestinal damage, indicated by the decrease in the villus-crypt ratio as aflatoxin concentration increased in the diet.2 It’s likely this caused interference with digestion and absorption of nutrients and contributed to the decreased average daily gain and bodyweight in ducklings fed the highest concentration of aflatoxin compared to the Calibrin-A control.1 Calibrin-A had a positive effect in the intestine by improving villus height and villus-crypt ratio in the duodenum and jejunum versus diets with no Calibrin-A.2

Analysis of liver enzymes indicated that serious liver damage also occurred in the high aflatoxin group. Creatine kinase, alanine transaminase and aspartate transaminase significantly decreased and alkaline phosphatase activity increased compared to the negative control clean diet, but the addition of Calibrin-A to the high aflatoxin diet neutralized these effects.1 All serum metabolites measured decreased in the high aflatoxin diet compared to clean diet; however, Calibrin-A improved serum metabolite concentrations.1

In the high aflatoxin group compared to the negative control clean diet, hepatoxicity was indicated by significantly decreased antioxidant defense systems — serum superoxide dismutase activity and serum and liver glutathione peroxidase activities — but they were improved with the addition of Calibrin-A to the diet.1 Serum and liver malondialdehyde concentration (an oxidative stress and liver damage marker) was also increased in the high aflatoxin group compared to the clean control, but Calibrin-A was able to prevent the increase.1

In these studies, feeding 0.1% Calibrin-A alleviated the aflatoxin-induced effects of reduced growth performance, increased mortality, liver damage, increased oxidative stress and impaired intestinal morphology of ducklings.

Calibrin-A Improves Aflatoxin-Challenged Broiler Performance

In a 21-day study at the University of Missouri, 320 day-old male broiler chicks (eight treatments, with eight replicate pens of five chicks per treatment) were fed diets with and without various concentrations of aflatoxin and Calibrin-A in the diet (Table 2).3

Table 2: Broiler chicks were assigned to one of eight treatments, with or without aflatoxin and with or without Calibrin-A.

Broilers challenged with aflatoxin (2 or 3 ppm) and fed either 0.25 or 0.5% Calibrin-A, had significantly greater body weight gain and feed intake compared to the aflatoxin-challenged controls (2 or 3 ppm aflatoxin; P < 0.05; Figure 1). At 2 ppm aflatoxin, both Calibrin-A concentrations also significantly reduced relative liver weight compared to the 2 ppm aflatoxin control (P < 0.05). No other groups were significantly different, although at 3 ppm aflatoxin, both Calibrin-A groups were numerically lower than the aflatoxin control (Figure 2).

This study confirmed that the addition of 0.5% Calibrin-A to the ration did not negatively affect broiler performance. Feed intake, body weight gain and feed conversion were not different between the negative control and 0.5% Calibrin-A, and neither were serum albumin, globulin, total protein, calcium or glucose concentrations.


Figure 1: Calibrin-A included at 0.25 and 0.5% had significantly greater body weight gain and feed intake compared to the mycotoxin control fed at either 2 ppm or 3 ppm (P < 0.05).

Figure 2: At 2 ppm aflatoxin, both Calibrin-A concentrations significantly reduced relative liver weight compared to the positive control (P < 0.05).


These studies show that Calibrin-A is a safe and effective solution for managing the toxic effects of aflatoxin in ducklings and broiler chicks — without interfering with nutrient utilization required for normal growth. For more details about these studies or to learn more about Calibrin-A, contact your local Amlan sales representative.



  1. Li, Y, Liu YH, Yang ZB, Wan XL and Chi F. The efficiency of clay enterosorbent to ameliorate the toxicity of aflatoxin B1 from contaminated corn (Zea mays) on hematology, serum biochemistry, and oxidative stress in ducklings. J Appl Poult Res. 2012; 21:806–815.
  2. Wan, XL, Yang ZB, Yang WR, Jiang SZ, Zhang GG, Johnston SL and Chi F. Toxicity of increasing aflatoxin B1 concentrations from contaminated corn with or without clay adsorbent supplementation in ducklings. Poult Sci. 2013; 92:1244–1253.
  3. Ledoux DR, Rottinghaus GE, Bermudez, AJ and Broomhead, J. Efficacy of the adsorbent Calibrin-A in ameliorating the toxic effects of aflatoxin in broiler chicks. Presented at International Poultry Scientific Forum, Atlanta, GA.


Calibrin®-Z Is Not Your Typical Biotoxin Binder

For decades, clays have been used globally as feed additives to help with feed flowability and pellet binding (e.g., Flo-Fre™ and Pel-Unite™). More recently, some international markets have also recognized the ability of mineral-based products to bind biotoxins in the intestinal tract of animals. However, there is a misconception in the industry that all clay minerals are the same and have the same biotoxin-binding abilities. The truth is that the natural properties of the minerals and how they’re processed have a significant impact on their effectiveness as a biotoxin enterosorbent.

Calibrin®-Z (available in select international markets) is an example of a mineral-based product that’s not your typical biotoxin-binding feed additive. Many mineral-based products focus on binding only one biotoxin (e.g., aflatoxin). However, the natural composition of Calibrin-Z and the proprietary processing technique that’s used to modify its surface chemistry allow it to bind a broad spectrum of biotoxins, including bacterial exotoxins and endotoxins and polar and nonpolar mycotoxins.

A Natural, Synergistic Mineral Mix

Calibrin-Z is a calcium bentonite that is primarily composed of two minerals — calcium montmorillonite and opal (amorphous silica lepispheres). Each of these minerals has specific physical properties and it’s how they’re naturally mixed together that gives Calibrin-Z part of its distinctive enterosorbent abilities (Figure 1).

Figure 1: A representation of the calcium montmorillonite layers intertwined with opal lepispheres.

The interwoven, naturally occurring calcium montmorillonite and opal lepispheres of Calibrin-Z create more biotoxin-binding opportunities than if they were used alone. It’s what makes the intimately mixed mineralogic nature of Calibrin-Z difficult to duplicate — the minerals grew and altered their properties for over 60 million years.

An Interconnected Pore Network

Calibrin-Z is very porous — it’s about 50 percent pore space consisting of nano, micro, meso and macro pore sizes. Opal lepispheres have their own micro pores and help to open up spaces within the calcium montmorillonite structure because they’re naturally formed amongst the calcium montmorillonite. This natural mixture of minerals gives Calibrin-Z a very low bulk density, high porosity and an extensive surface area of around 450,000 m2/kg — an area equivalent to around 50 average-sized soccer stadiums. A phenomenal amount of chemistry can occur on this expansive surface area.

In order for Calibrin-Z to adsorb biotoxins that are in intestinal fluid, the fluid must first be absorbed into its pores. The pores of Calibrin-Z are an interconnected network that allows intestinal fluid to enter them, where the biotoxins can then interact or bind with the pore surface. Unlike other clays, the porosity of Calibrin-Z creates an open construct on a microscopic scale and a more inviting place for intestinal fluids to be absorbed into.

A Hydrophobic Surface Improves Biotoxin Binding

When the calcium bentonite in Calibrin-Z is mined, the capillaries and channels of its pores are naturally filled with water. Our proprietary thermal processing technique removes most of this water and also modifies the surface of the minerals to allow the pore surfaces to become more hydrophobic (Figure 2). This is a significant point of difference for Calibrin-Z because certain biotoxins (e.g., nonpolar mycotoxins and bacterial toxins) have a greater affinity for hydrophobic surfaces than hydrophilic surfaces. Other mineral-based products that are more hydrophilic than hydrophobic won’t be able to adsorb the broad spectrum of bacterial and fungal toxins that Calibrin-Z can bind.

Figure 2: Thermal processing removes water and changes the surface chemistry to create biotoxin-binding Calibrin-Z.

Safe, Irreversible Thermal Processing

Our proprietary thermal processing heats the surface of Calibrin-Z, changing its surface chemistry. These thermochemical changes are irreversible — they can’t be undone even after being exposed to the harsh environment of the animal’s gut.

Some companies activate the surface of their minerals with harsh chemicals instead of heat. Chemical activation of minerals does not create an irreversible bond, so theoretically these chemicals could be stripped off in the animal’s gut. This is important because the in vitro biotoxin-binding efficacy of a mineral-based product activated with chemicals may not be replicated in the real world. In the animal’s gut, the minerals are exposed to strong acids, enzymes and other substances that can change their properties and biotoxin enterosorbent abilities. Thermal processing is a safer, ecofriendly alternative to chemical activation of minerals that is permanent.

Vertical Integration Ensures Consistency

Quality and reliability of supply is another differentiator for Calibrin-Z. Amlan is the animal health business of Oil-Dri® Corporation of America, a leading global manufacturer and marketer of sorbent minerals. Vertical integration allows Oil-Dri and Amlan to control every step of the mineral production process to reliably deliver safe, high-quality and efficacious mineral-based products.

If needed, we can trace the minerals in our products from the feed bag, through the supply chain, through the processing chain in our plants back to where we mined the calcium bentonite. This includes the original core drill hole that was used to extract and pre-qualify the raw material to ensure it met the physical and absorptive quality profile. We mine our calcium bentonite from a single mine and from a specific location within our mine that has hundreds of years of reserves — giving our customers confidence in the availability, consistency, quality and efficacy of our minerals every time they buy a new batch of products.


The biotoxin-binding abilities of Calibrin-Z are quite different compared to most other calcium bentonite-based biotoxin binders. That’s why it’s important to look beyond the ingredient list on the feed bag and review the in vivo research that proves a feed additive can improve the intestinal health and performance of production animals. To read our in vivo Calibrin-Z research, visit our Research page or contact your local Amlan sales representative.


Natural Mineral-Based Neutralization of E. coli Toxins

Bacterial pathogens such as Escherichia coli (E. coli) are a leading cause of enteric disease and economic loss for animal protein producers. Found readily in the environment of poultry and swine, E. coli can produce powerful toxins (exotoxins and endotoxins, see Figure 1) that damage the integrity and function of the intestinal barrier. These toxins cause inflammation, diarrhea and malabsorption that result in less gain, higher feed conversion and reduced performance. However, multiple studies have confirmed that natural mineral-based feed additives can provide an antibiotic-free method of reducing the negative production effects of E. coli toxins.

Figure 1: E. coli exotoxins are produced by the cell and secreted into the environment. Endotoxin (lipopolysaccharide, LPS) is released from the cell wall of E. coli or other gram-negative bacteria after the bacteria dies.


Powerful Toxins

Multiple E. coli strains produce potent exotoxins as part of their virulence package. These powerful toxins are major causes of enteric disease, particularly in neonatal and recently weaned animals. The toxins damage the intestinal lining, resulting in diarrhea, dehydration and even death.

Heat-Labile Enterotoxin

Enterotoxigenic E. coli (ETEC) — a common cause of porcine post-weaning diarrhea (PWD) and a source of diarrhea in poultry — produces the enterotoxin heat-labile toxin (LT). To exert its effects, LT is internalized by binding to surface receptors on the host intestinal mucosa.1 Once inside the cell, LT alters the electrolyte and water balance of the cell, which leads to diarrhea.1 A survey of U.S. ETEC isolates linked to porcine PWD showed that 57.7% had the heat-labile toxin gene.2

Shiga-like Toxin

E. coli types that produce shiga-like toxins are referred to as shiga toxin-producing E. coli (STEC). There are two types of shiga-like toxins (Stx1 and Stx2), and both have multiple subtypes. The mode of action for shiga-like toxin is inhibition of host cell protein synthesis that results in cell death.3 Edema disease (E. coli enterotoxemia) in swine is caused by shiga-like toxin (Stx2) subtype e.4

E. coli Endotoxin

The outer membrane of gram-negative bacteria such as E. coli or Salmonella contains lipopolysaccharides (LPS) which protect the bacteria from the environment. When the bacteria die naturally or as a result of using antibacterial products (e.g., antibiotics, specialized feed additives), endotoxin is released from the cell wall. Endotoxin causes specialized immune cells in the animal’s body to release high levels of cytokines that result in elevated temperature, weakness, tissue necrosis and, at high levels, death.

Management Programs to Control E. coli

With the global push toward antibiotic-free production, maintaining a healthy intestinal environment to reduce the risk of disease from E. coli now requires effective management strategies. Natural feed additives, tight biosecurity, effective sanitation, and good husbandry practices that protect the intestinal environment can all help control the risk of E. coli outbreaks.

Natural Mineral-Based Feed Additives Neutralize E. coli Toxins

The backbone of Amlan’s scientifically proven products is our natural mineral that undergoes proprietary thermal processing to optimize the efficacy of each product. Varium® (available in select international markets) for poultry and NeoPrime® (available in select international markets) for swine are species-specific formulations of three synergistic ingredients that improve feed conversion and intestinal integrity in poultry and maximize post-weaning performance in swine. Calibrin®-Z (available in select international markets) is a low-dose biotoxin binder for all species that actively defends the intestinal barrier against a broad spectrum of toxins produced by bacteria and fungi.

Adsorption of E. coli toxins by Varium and Calibrin-Z

The ability of Varium and Calibrin-Z to adsorb E. coli toxins was assessed in vitro by the United States Department of Agriculture Agricultural Research Service (USDA-ARS), Beltsville, MD. Calibrin-Z or Varium was added separately to each toxin, incubated to allow the toxin to bind, and then centrifuged to separate the free and bound toxins. The USDA researchers had developed a specific ELISA for heat-labile and shiga-like toxins, which was used to analyze the supernatant to quantify the presence of unbound toxins.

Varium and Calibrin-Z were both effective at neutralizing both exotoxins (Figures 2 and 3) and endotoxins. (Figure 4).


Figure 2: Varium neutralized E. coli exotoxins in vitro. Source: USDA-ARS, Beltsville, MD.

Figure 3: Calibrin-Z adsorbed E. coli exotoxins in vitro. Source: USDA-ARS, Beltsville, MD.

Figure 4: Varium and Calibrin-Z adsorbed E. coli endotoxin (LPS) in vitro. Calibrin-Z source: Gibraltar Laboratories, Fairfield, NJ. Varium source: USDA-ARS, Beltsville, MD.

NeoPrime Reduces Plasma Endotoxin Concentration

When endotoxin is released from dead gram-negative bacteria, it can damage the intestinal epithelial cells. Weakness in the intestinal barrier can lead to “leaky gut syndrome,” which allows the endotoxin to enter the blood stream. NeoPrime was assessed for its ability to strengthen intestinal integrity by analyzing the blood plasma endotoxin concentration of weanling pigs. Compared to a control that contained two antibiotics and zinc oxide, NeoPrime in multiple diet formulations reduced plasma endotoxin concentration from E. coli or other gram-negative bacteria (Figure 5).

Figure 5: Feeding NeoPrime decreased plasma endotoxins, indicating improved gut barrier function Source: Hunan Agricultural University, Hunan, China.


When to Use Our Mineral-Based Feed Additives

The research presented here supports in vivo studies showing that supplementation with Varium, NeoPrime or Calibrin-Z is an effective approach to reducing the effects of enteric disease in poultry and swine. To learn more about these natural mineral-based control methods for improving intestinal health and production efficiency, visit or contact us at




  1. Duan Q, Xia P, Nandre R, Zhang W, Zhu G. Review of newly identified functions associated with the heat-labile toxin of enterotoxigenic Escherichia coli. Front Cell Infect Microbiol. 2019.
  2. Zhang W, Zhao M, Ruesch L, Omot A, Francis D. Prevalence of virulence genes in Escherichia coli strains recently isolated from young pigs with diarrhea in the US. Veterinary Microbiology. 2007;123:145–152.
  3. Melton-Celsa A. Shiga toxin (Stx) classification, structure, and function. Microbiol Spec. 2014;2.
  4. Baldo V, Salogni C, Giovannini S, D’Incau M, Boniotti MB, Birbes L, Pitozzi A, Formenti N, Grassi A, Pasquali P, Alborali GL. Pathogenicity of shiga toxin type 2e Escherichia coli in pig colibacillosis. Front Vet Sci 2020 Sep 18;7: n.p. doi: 10.3389/fvets.2020.545818.