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Binding Bacterial and Fungal Toxins

Calibrin-Z logo with chickens in background.

Source: Poultry World, February 25, 2022

One of the biggest challenges for the poultry industry, especially after the removal of in-feed antibiotics, is bacterial enteritis, and in particular, necrotic enteritis caused by pathogenic Clostridium perfringens. Add production losses associated with mycotoxin-contaminated feed, and you have two industry challenges that need a natural control solution.

Calibrin-Z from Amlan International provides natural protection against field challenges, optimizing animal performance and boosting your bottom line. It is a highly efficacious, mineral-based feed additive that adsorbs a broad spectrum of bacterial toxins and mycotoxins in the intestinal tract of chickens. Unlike comparable products, Calibrin-Z binds both polar and nonpolar mycotoxins as well as bacterial toxins like alpha-toxin and NetB toxin that are produced by C. perfringens.

Read more here.

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Exploring Necrotic Enteritis: Cause, Effects and Solutions

Computer-generated illustration of necrotic enteritis.

Necrotic enteritis is a common infectious disease that costs the poultry industry billions of dollars each year. With more and more poultry production systems moving toward the global objective of reduced in-feed use of antibiotics for growth promotion, maintaining a healthy intestinal environment now relies more heavily on other management methods to reduce the risk of necrotic enteritis development. Keep reading to learn more about the origins of necrotic enteritis, the relationship between necrotic enteritis and coccidiosis, and management of the disease risk in ABF production systems.

C. perfringens: The Cause of Necrotic Enteritis

Necrotic enteritis is caused by Clostridium perfringens types A and C: gram-positive, spore-forming anerobic bacteria. C. perfringens can be found throughout the poultry house environment and has spores which, under the right environmental conditions, can survive for long periods outside the bird. The normal microbiota of the bird contains C. perfringens, so its presence alone doesn’t necessarily indicate an issue. Instead, disease occurs when predisposing conditions in the bird cause overgrowth of the pathogen. There are multiple factors that can contribute to C. perfringens overgrowth including diet changes, immune status and stress, intestinal pathophysiology and concurrent infection with coccidiosis.

Multiple exotoxins can be produced by C. perfringens, including alpha-toxin and necrotic enteritis toxin B-like toxin (NetB). Alpha-toxin is cytotoxic to endothelial cells, red blood cells, white blood cells and platelets, whereas NetB toxin forms pores in cell membranes that allows electrolytes to rupture cells, causing cell death and necrotic lesions in the small intestinal mucosa. These two toxins are known to have a role in necrotic enteritis development.

Impaired Nutrient Adsorption

In healthy birds, the epithelium and mucus layer of the intestine form a selective barrier between internal tissues and the external environment (the intestinal lumen). The natural barrier allows nutrients through the intestinal wall but not pathogens and their biotoxins. When there is an overgrowth of C. perfringens, there is a breakdown of the defensive barrier. The damage to the intestinal wall hinders nutrient digestion and absorption and can allow toxins to enter the circulatory system.

Necrotic enteritis can present as either clinical or subclinical disease. The clinical form is characterized by high mortality, whereas subclinical disease causes decreased weight gain and increased feed conversion ratio (FCR). The increased FCR is due to the reduced nutrient digestibility and adsorption, resulting in compensatory feed intake.

Coccidiosis Can Increase Necrotic Enteritis Incidence

Coccidiosis is another common disease in poultry that causes a significant negative economic impact on production. It is an enteric disease caused by parasitic protozoa in the Eimeria genus. Part of the Eimeria life cycle includes invading intestinal epithelial cells, damaging the intestinal wall barrier. The damage that coccidia cause to intestinal epithelial cells promotes invasion by C. perfringens (as well as other pathogens), increasing the incidence of necrotic enteritis.

Necrotic Enteritis in Antibiotic-Free Production

Managing necrotic enteritis in an ABF production system requires detailed strategies including the use of vaccinations, diagnostic tools and natural feed additives. In some production systems, ionophores used to control coccidiosis may also be eliminated, which further heightens the importance of other management practices that keep birds free of diseases like coccidiosis and necrotic enteritis.

Management practices, such as biosecurity, sanitation, water quality and air quality, also require increased planning and monitoring in ABF systems. Reduced ventilation, increased litter moisture and poor husbandry can increase the incidence of necrotic enteritis. The quality and sanitation of eggs in the hatchery also needs to be managed well in ABF systems, so that birds have a healthy start to life.

Necrotic Enteritis Control Solutions

Along with management best practices, natural feed additives such as Varium® and Calibrin®-Z are available to help reduce the incidence of necrotic enteritis. Varium, a natural patented mineral-based feed formulation, can reduce challenges from pathogenic bacteria and their toxins, strengthen the intestinal barrier and activate the innate immune system to naturally defend against disease. In high challenge environments, Calibrin-Z, a broad spectrum biotoxin control feed additive, can be used on top of Varium to reduce the level of pathogens in the intestines that cause mortality, wet droppings, reduced feed efficiency and damage the integrity of the intestine.

Necrotic enteritis is a challenging disease for poultry producers, particularly those using an ABF production system. However, with best practice management strategies and inclusion of feed additives that promote intestinal health and function, poultry flock health can be improved, thereby reducing the risk of disease and maximizing production efficiency. To learn more about necrotic enteritis, keep checking the Education Center for other posts in the necrotic enteritis series.  


Chi, F. A Viable Adjunct or Alternative to Antibiotics: Meta-Analysis of Broiler Research Shows Natural Growth Promoter Delivers Feed Efficiency Equal to Antibiotics, Amlan International.

An Antivirulence Approach to Controlling Bacteria: AGP Alternatives in Development at the Richard M. Jaffee Center for Applied Microbiology

Computer-generated image of bacteria.

The rise of multi-drug-resistant and mutant bacteria, which don’t respond to antibiotic treatment, is a concerning trend across the world. With the use of antibiotic growth promoters (AGPs) restricted in an increasing number of countries, livestock and poultry industries are looking for antibiotic-free alternatives that keep animals and birds healthy and productive. Amlan International’s Richard M. Jaffee Center for Applied Microbiology is developing industry-leading technology that helps livestock and poultry producers achieve peak antibiotic-free-performance.

Discovering new methods to combat the prevalence of resistant and mutant pathogens is just one of the topics scientists at the Richard M. Jaffee Center for Applied Microbiology are investigating. The research team at Amlan’s new state-of-the-art lab are using an antivirulence approach to control pathogens, which has also been a research focus for developing new antimicrobial drugs for humans.

Novel Alternative to Antibiotics

Unlike conventional antibiotics, antivirulence approach targets bacterial virulence factors and is aimed at disarming pathogens and modifying their behaviors by making them less harmful (less virulent) to the host. The likelihood for multi-drug-resistant and mutant bacteria is much less using this approach.

There are many options for controlling pathogens using an antivirulence approach. These antivirulence targets include:

  • Toxins
  • Secretion systems
  • Quorum sensing
  • Host-pathogen signaling
  • Adhesins
  • Biofilm formation
  • Siderophores
  • Immune evasion

Previous research at Amlan International showed exciting promise in the quorum sensing area. The Richard M. Jaffee Center for Applied Microbiology continues to conduct quorum sensing research as one component of the overall antivirulence approach to improving livestock and poultry health and production.

Bacteria Can Communicate

Quorum sensing is a communication system between bacterial cells that allows bacteria to regulate their activity in response to stimuli. This communication system involves bacteria releasing biochemicals into the environment which accumulate in the surrounding area until reaching a critical threshold concentration1. The biochemicals then bind to receptors on the bacteria, signaling gene expression.

Quorum sensing can control many functions in bacteria2 including:

  • Bacterial population
  • Virulent gene expression
  • Bioluminescence
  • Pigment generation
  • Bacterial swarming
  • Biofilm formation

Quorum Quenching Silences Bacteria

Quorum quenching is an approach which can disrupt the quorum sensing system of pathogenic bacteria, preventing cell-to-cell communication and the expression of virulence genes that could lead to their infection in the host. Quorum quenching products should reduce the chance of antibiotic resistance, since they are modifying bacteria behavior rather than killing them.

Calibrin®-Z Disrupts Quorum Sensing

Evidence of quorum quenching by Amlan’s Calibrin®‑Z biotoxin control product was published in the Journal of Agricultural and Food Chemistry. The research team at the Richard M. Jaffee Center for Applied Microbiology are continuing to build on these positive results and further quorum sensing research.

The previous study illustrated that in vitro, quorum sensing molecules can be separated out by adsorption or catalytically broken down into small fragments by Calibrin-Z. By reducing the concentration of quorum sensing biochemicals, the products can potentially disrupt the ability of pathogenic bacteria to produce toxins or reduce their virulence, since these functions are controlled through quorum sensing. 

Additionally, due to their stronger acidity, greater pore volume and greater surface area, in the same study Calibrin-Z performed better then silica, illite and kaolinite as catalysts/adsorbents in quorum sensing molecule removal.

When incubated with Vibrio harveyi, a bacterium that exhibits bioluminescence controlled via quorum signaling, Calibrin-Z reduced bacterial luminescence by 55% (from the area under the curve; Figure 1). While this demonstrated that interference in quorum sensing occurred, the bacterial numbers were not impacted, indicating the reduction in bioluminescence was achieved through quorum quenching and not by killing bacteria.

Bacterial luminescence graph.

Figure 1: Bacterial luminescence from a Vibrio harveyi culture treated with different concentrations of Calibrin-Z. Vibrio parahaemolyticus (Vp) was used as a non-luminescent negative control. At 10 mg/mL, Calibrin-Z reduced bacterial luminescence by 55% (area under the curve).

Quorum sensing is just one of the virulence factors being investigated by the research team at the Richard M. Jaffee Center for Applied Microbiology. Similarly, the antivirulence approach is just one of the next-gen technologies that is being developed in the lab to maximize livestock and poultry producer’s profits by improving the health and productivity of their flock or herd.

Stay tuned for more information about the innovative research conducted inside the Richard M. Jaffee Center for Applied Microbiology.


  1. Naik, S.P., Scholin, J., Ching, S., Chi, F. and Herpfer, M. (2018). Quorum Sensing Disruption in Vibrio harveyi Bacteria by Clay Materials. Journal of Agricultural and Food Chemistry, 66 (1), 40-44. DOI: 10.1021/acs.jafc.7b03918
  2. Williams, P. (2007) Quorum Sensing, Communication and Cross Kingdom Signalling in the Bacterial World. Microbiology, 153 (12), 3923−3938. DOI: 10.1099/mic.0.2007/012856-0

Safeguard Livestock Health and Productivity with On-Site Mycotoxin Diagnostics and Product Dosing Guide

Bio Insights Mycotoxin Diagnostics by Amlan International

Mycotoxins are a natural product of fungal metabolism. However, when they end up in livestock feed, they are of critical concern for producers worldwide because of their detrimental impact on health and productivity: they can reduce feed intake and efficiency, decrease weight gain, damage internal organs such as the liver and kidneys and increase mortality.1

Despite several mycotoxin binder products in the market, mycotoxins continue to be a problem; a 2013 global mycotoxin survey revealed that 81% of grain and feed samples analyzed contained at least one mycotoxin.2

Previously, the only way to determine if feed was contaminated with mycotoxins was to send samples to outside laboratories for analysis. With BioInsights Mycotoxin Diagnostic Services, producers can test feed ingredients on-site and accurately identify mycotoxins in minutes.  As a result, producers can eliminate the cost and delay of sending samples to outside laboratories for testing and optimally dose Amlan International’s biotoxin control product, Calibrin-Z.

Using its extensive mineral chemistry experience, expertise in animal science and state-of-the-art microbiology lab, coupled with a partnership with a diagnostic leader in the industry, Amlan International has developed this program to help its partners raise animals safely and efficiently by providing biological insights to prevent production losses.

Minimize production losses with timely mycotoxin diagnostic results

To bring producers the first diagnostic system for fast and accurate on-site mycotoxin detection, Amlan International partnered with Envirologix, the world’s most trusted name for quantitative GMO testing at the point of grain delivery. 

Using a standard PC computer platform, the Envirologix QuickScan II system works with mycotoxin test kits to provide rapid, accurate quantitative results. In under 10 minutes, the system extracts, incubates and identifies mycotoxins in feed. Producers can then use Amlan International’s dose calculator to determine the optimal dose of adsorbents, such as Calibrin®-Z, to protect their animals.

The dangers of multiple mycotoxins in poultry feed

In many cases, feed may be contaminated with multiple mycotoxins. Together, they can have a synergistic, or additive effect, meaning the negative effects of one mycotoxin are amplified by the interaction with another, making their combined effect greater than the sum of their individual effects. Producers are encouraged to consider the total toxin load when dosing mycotoxin adsorbents.


  1. Filazi A, Yurdakok-Dikmen B, Kuzukiran O, Sireli UT. Mycotoxins in Poultry 2017; DOI: 10.5772/66302. Available at Accessed August 6, 2020.
  2. Murugesan GR, Ledoux DR, Naehrer K, et al. Prevalence and effects of mycotoxins on poultry health and performance, and recent development in mycotoxin counteracting strategies. Poult Sci 2015;94(6):1298-1315.