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Calibrin®-Z: An In-Depth Look at the Speed and Superior Binding Efficacy

An Urgent Need for Antibiotic Alternatives

Pseudomonas aeruginosa is a bacterial species, capable of surviving in a variety of animal and plant hosts and causing opportunistic infections. Its resistance to multiple antibiotics and disinfectants makes it a significant concern in modern medicine and animal production facilities. The World Health Organization lists carbapenem resistant P. aeruginosa as one of the top three pathogens in which new antibiotics or alternative treatments are critically needed. Aside from human medicine, livestock and companion animals are known hosts for serious infections from P. aeruginosa. Notable infections include mastitis in cattle, respiratory diseases in poultry, ear infections in dogs and reproductive infections in horses (Table 1). P. aeruginosa infections in livestock have a significant economic impact due to factors like increased morbidity and mortality, reduced production yields, higher veterinary costs for treatment and the need for stricter biosecurity measures to prevent spread. Given its pathogenic potential and resistance to antibiotics, P. aeruginosa infections in livestock require effective preventive measures and targeted therapeutic strategies to minimize economic losses and animal health impacts. 

Table 1. List of diseases caused by P. aeruginosa in various hosts

Pyocyanin, a Key Virulence Factor  

One of the crucial toxins produced by P. aeruginosa is pyocyanin. It is a phenazine compound known for its role as a virulence factor, contributing to the pathogenicity of the bacteria. Pyocyanin generates reactive oxygen species (ROS), which can damage host cells by disrupting cellular functions, impairing immune responses, and inducing oxidative stress. It also interferes with cellular respiration and signaling pathways. Pyocyanin is notable for its characteristic color and is often used as a diagnostic marker for P. aeruginosa infections. Its ability to modulate the host immune system and disrupt cellular functions makes it a key target for research on antibacterial strategies and therapeutic interventions.

Fig. 1 | Pyocyanin is a nitrogen-containing aromatic metabolite. The color of pyocyanin is blue-greenish at the neutral and alkaline pH levels and that changes to purple-red in acidic conditions. It has a low molecular weight, which enables easy biological membrane permeation.

A proven toxin binder 

Calibrin-Z is backed by over 15 years of research, proven to bind bacterial pathogens and their toxins, as well as mycotoxins, protecting production animals from a broad spectrum of biotoxins that reduce performance and cause morbidity or mortality. A one-ingredient feed additive, Calibrin-Z is made from our selectively sourced calcium montmorillonite with opal-CT lepispheres that undergoes proprietary thermal processing to promote the binding of multiple biotoxins in the intestine. Calibrin-Z’s ability to rapidly bind this broad spectrum of toxins sets it apart from others in its class. 

During research to determine if Calibrin-Z would rapidly target pyocyanin, two pyocyanin solutions were used. As shown in Figure 2, the pyocyanin solution is purple/red at pH 3 and blue at pH 7. Granules of Calibrin-Z were added into each solution. Then the samples were mixed briefly by vortexing. When minerals settled on the bottom, the pyocyanin solution looked clear suggesting the removal of pyocyanin from water solution. The fact that Calibrin-Z neutralized pyocyanin demonstrates its speed and effectiveness toward harmful toxins.  

Commercial Farms Recognize the Proven Benefits of Calibrin-Z 

Calibrin-Z offers the critical advantage of rapid toxin binding, helping prevent harmful biotoxins from being absorbed into the bloodstream—something that can happen within minutes of ingestion. Once toxins enter the bloodstream, they can causesystemic damage and disease, significantly reducing animal performance while also compromising the safety of meat, milk, and eggs. By using a proven toxin binder like Calibrin-Z, producers can help protect both animal health and food security. Backed by numerous commercial case studies, Calibrin-Z consistently delivers results,  making it a reliable solution for both poultry and livestock production. 

To begin your own Calibrin-Z trial, or for more information about the benefits of using Calibrin-Z for biotoxin control, contact us at info@amlan.com. 

References: 

Wang, D. and H. Xue. 2021. Clay mineral-based treatments in Pseudomonas aeruginosa infection control. United States Patent Application. Application No.: 17/504,338 

 

NeutraPath® Effectively Inhibits Salmonella Colonization in Poultry

Salmonella is one of the most common food-borne pathogens. The CDC estimates that Salmonella causes about 1.35 million illnesses in the United States each year, resulting in 26,500 hospitalizations annually. Previously, subtherapeutic levels of antibiotics used for growth promotion could control enteric pathogens like Salmonella. However, concerns about antibiotic resistance have led to regulations and restrictions on the use of antibiotics in animal feed. As a result, the demand for antibiotic-free alternatives that control enteric pathogens has increased dramatically.  

 To meet this growing need, Amlan International offers innovative solutions that support poultry health and performance without the use of antibiotics.   

 NeutraPath®, available in select international markets, optimizes poultry production efficiency and economic performance through a proprietary blend of essential oils, medium-chain fatty acids, and Amlan’s thermally processed calcium montmorillonite. This synergistic formula works through multiple modes of action to improve intestinal health, boost feed efficiency, and reduce mortality in disease challenged birds. 

NeutraPath: Proven to Inhibit Salmonella and Reduce Colonization

A study at the University of Arkansas evaluated  NeutraPath’s ability to inhibit Salmonella enterica serovar Typhimurium PHL2020 isolate (ST-PHL2020) intestinal colonization in broiler chickens. The research demonstrated that NeutraPath effectively inhibits ST-PHL2020 colonization in various parts of the upper gastrointestinal tract using an in vitro model designed to simulates conditions of a live digestive environment.   

 The in vitro digestion model consisted of three compartments that were simulated to match the pH, enzymatic composition, and temperature of the crop, proventriculus, and intestinal section of the chicken gastrointestinal tract.   

 The lowest concentration of NeutraPath used in this model (0.25%) reduced ST-PHL2020 colonization in the proventriculus and intestinal compartments compared to the control. An increase of NeutraPath concentration led to increased inhibition of ST-PHL2020 colonization. The 0.5% treatment reduced colonization in the crop and there was no detectable colonization in the proventriculus and intestinal compartments.   

The in vivo section of this study revealed that a 0.25% supplementation of NeutraPath resulted in a 2-log reduction of bacterial load in the cecal tonsils compared to the control. This dosage of NeutraPath also reduced total prevalence of Salmonella in the ceca to 58% compared to 100% in the control. The findings from this study suggest that the unique blend of Amlan’s mineral, select essential oils and medium chain fatty acids inhibited Salmonella spp. in vitro and decreased Salmonella spp. populations within the chicken ceca in vivo.

Downregulation of Salmonella Virulence Genes

In addition to the inhibitory effect on Salmonella growth, NeutraPath also displayed the ability to modulate virulence gene expression. ST-PHL2020 was grown in LB culture media with or without supplementation of 1mg/mL of NeutraPath for 12 hours.  

 After incubation, RNA was extracted, reverse-transcribed, and then subjected to qPCR to analyze the expression of several Salmonella virulence genes. The expression of hilA and invF in samples treated with 0.5g/mL of NeutraPath was downregulated 1.71 and 10.71-fold respectively when compared to the control. The hilA and invF genes are necessary for the assembly of the TTSS (Type III secretion system) within Salmonella that is responsible for delivering effector proteins into the cytoplasm of host cells to promote virulence and colonization. NeutraPath also downregulated the expression of sopB, sopE, and sipA compared to the control. These genes encode effector proteins that stimulate inflammation and enhance the efficiency of host cell invasion.

These findings demonstrate NeutraPath’s ability to inhibit bacterial colonization in the chicken gastrointestinal tract and to neutralize important virulence factors. The results suggest that NeutraPath can be a viable alternative to antibiotics for controlling enteric pathogens in poultry production. To learn more about NeutraPath, visit amlan.com or connect with a local representative here. 

References:  

Eichelberg, K., & GaláN, J. E. (1999). Differential regulation of Salmonella typhimurium Type III secreted proteins by Pathogenicity Island 1 (SPI-1)-Encoded Transcriptional Activators INVF and HILA. Infection and Immunity, 67(8), 4099–4105. https://doi.org/10.1128/iai.67.8.4099-4105.1999  

 Xue, H., Wang, D., Hargis, B., & Tellez-Isaias, G. (2022). Research Note: Virulence gene downregulation and reduced intestinal colonization of Salmonella enterica serovar Typhimurium PHL2020 isolate in broilers by a natural antimicrobial (NeutraPathTM). Poultry Science, 101(6), 101822.  

Centers for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious Diseases (NCEZID), Division of Foodborne, Waterborne, and Environmental Diseases (DFWED) 

 

Billions with a B: The Global Cost of Coccidiosis in Poultry

Estimates of the global cost of coccidiosis in poultry vary widely, but most place the figure in the tens of billions of U.S. dollars. Coccidiosis is characterized by symptoms such as diarrhea, intestinal lesions, morbidity, and mortality, along with decreased weight gain and feed efficiency.   The intestinal damage caused by the disease can lead to secondary issues such as necrotic enteritis, which has become more prevalent as poultry producers reduce antibiotic use.  The majority of losses stem from reduced production due to subclinical coccidiosis.  The lack of overt disease emphasizes the importance of vigilance and proactive management systems.

To combat the impact of coccidiosis in poultry, producers rely on a range of methods, each offering unique benefits and presenting specific limitations.  Traditional control methods include the use of vaccines, ionophores, and chemical coccidostats.  However, natural alternatives for managing coccidiosis have been developed through the years, including the use of phytogenic compounds.

A Sustainable Natural Alternative

Phylox® (available from Amlan International in select international markets), a natural feed additive featuring a select blend of bioactive phytochemicals was researched to determine its effects on broilers challenged with multiple species of Eimeria. Phylox is designed to interrupt the complex Eimeria lifecycle at several key points, preventing damage to the host and reducing the risk of growing resistance. It works by compromising the sporozoites’ cell membrane, inhibiting oocyst sporulation, enhancing anticoccidial immunity, and supplying essential energy to the intestinal cells. By targeting all three stages of the coccidia lifecycle, Phylox helps protect intestinal integrity and support overall bird health. 

Research shows that Phylox decreases the effects of coccidiosis on gut health and improves bird performance. Two experiments that lasted 28 days each were used to study its effects on young broilers during an Eimeria challenge. Both studies used a multi-species coccidia challenge that included giving an oral dose of E. maxima (50,000 oocysts), E. acervulina (100,000 oocysts), and E. tenella (75,000 oocysts) to each bird on the 14th day of the experiment. Six days post-challenge five birds per pen (eight pens per treatment) were euthanized so that intestines could be inspected for coccidial lesions. The Johnson and Reid (1970) scoring method was used to assess the severity of intestinal lesions caused by coccidiosis, with a score from 0 (no visible lesions) to 4 (severe, widespread lesions). No coccidia vaccine or medication was used in the first study, which evaluated the effect of Phylox on both unchallenged and challenged broilers. The second study was conducted to compare Phylox to drugs currently used to control coccidiosis in the field.

Feeding Phylox improved gut health in broilers challenged with multiple Eimeria species. This was evidenced by lower coccidia lesion scores in birds fed Phylox compared to untreated, challenged birds (Study 1), with scores decreasing from 2.43 to 1.85. Lower lesion scores, indicating improved gut health, are expected to result in better feed efficiency. Feeding Phylox to birds challenged with multiple Eimeria species improved feed conversion, both during the critical period following the challenge and overall. Additionally, no negative effects were observed in unchallenged birds fed Phylox.

In Study Two, feeding Phylox, salinomycin, or Nicarbazin equally reduced lesion scores compared to challenged but untreated birds. The resulting improvement in gut health led to improved feed conversion ratios for all treatments. Each of the three coccidiosis treatments produced similar results, with improvement in feed conversion observed both during the post-challenge period and throughout the overall experimental period. This research demonstrates that feeding Phylox to Eimeria-challenged broilers mitigated the effects of coccidiosis, with outcomes comparable to those of commonly used treatments.

Immune Function

The gut of any animal faces constant challenges. Broilers continuously ingest bacteria, parasites, toxins, and other harmful substances along with their feed and water. The intestinal wall acts as a crucial barrier, preventing these pathogens from entering the body. This is why the majority of a chicken’s immune tissue is located in the gastrointestinal tract.

A third experiment examined the effects of coccidia and Phylox on immunity and gut microbiota. The research showed that Phylox helped preserve immune homeostasis by enhancing the anticoccidial immune response during coccidial challenge. Specifically, Phylox preserved the function of protective mucosal immunity by promoting a non-terminally differentiated helper T-cell subset (CD8-CD28+). This helps ensure a competent T-cell response during pathogen challenges, such as the Eimeria challenge used in this experiment.

Additionally, Phylox reduced Eimeria challenge-associated upregulation of cecal IL-10. IL-10 is an anti-inflammatory cytokine with potent immunosuppressive effects. Intracellular parasites like coccidia can use the immunosuppressive properties of IL-10 to help them survive in a hostile environment. When coccidia causes an increase in IL-10 during a challenge, as seen in this experiment, it results in a decreased immune response just when the animal needs the immune response to help fight the disease. Phylox fed broilers showed suppression of IL-10 during the coccidia infection, allowing a proper and effective immune response to occur. This was shown by decreased cecal IL-10 mRNA when Phylox was fed.

Summary

To mitigate losses associated with coccidiosis in poultry, innovative solutions are essential to complement existing vaccines and drugs. Producers require new, consumer-accepted alternatives to effectively manage coccidiosis on their farms. Phylox serves as a valuable addition to the available tools, helping to address this persistent challenge and support poultry health and productivity. To learn more about Phylox, or to begin a field trial, contact your local Amlan representative here.

Dr. Aldo Rossi Talks Speed of Binding with Feedstuffs 365

At IPPE 2025, Dr. Aldo Rossi sat down with Ann Hess of Feedstuffs to discuss the critical role of binding speed in biotoxin control. Many toxins in contaminated feed can be absorbed into a bird’s digestive tract within just 30 minutes of consumption. This makes the speed at which a feed additive acts essential for protecting animal health.

Amlan’s Calibrin®-Z delivers rapid, broad-spectrum toxin binding, helping safeguard poultry health and support optimal productivity. Learn how its fast-acting, comprehensive binding capabilities are advancing animal health and improving performance for producers worldwide. To watch the interview, click here.

Consumer demand driving natural feed solutions

Poultry barn with Amlan logo text graphic.

Source: Amie Simpson, Brownfield Ag NewsJanuary 28, 2022

“Increasing regulation and strong consumer demand are creating a growing market for natural animal protein production solutions,” commented Dr. Wade Robey, our vice president of marketing and product development, in a recent interview with Amie Simpson of Brownfield Ag News. Wade also discussed Amlan’s introduction of natural mineral-based food additives to the US market and the growth opportunities this presents for Amlan in 2022. Read more here.

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