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Calibrin®-Z: A Solution for Performance-Degrading Challenges Caused by Lethal Toxins

August 8, 2023
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Normally when we think of problems caused by bacteria, we think of the diseases and the damage that they do when they are alive. We believe if we kill the bacteria, we will stop the disease that they cause to ourselves or our livestock. Unfortunately, the death or destruction of some bacteria does not end the damage that they can cause.

Gram negative bacteria such as Escherichia coli have a component of the outer membrane of the cell wall called lipopolysaccharides (LPS). Lipopolysaccharides, also referred to as endotoxins, are highly toxic and are released when the cell dies, is damaged, or destroyed. Lipopolysaccharides range in size from 10 thousand to 1 million daltons. Because of their small size they can easily enter the bloodstream through a damaged or “leaky” intestine. When this happens endotoxins can be very toxic to humans and animals. How toxic depends first on the species of animal and the amount of endotoxins that enter the bloodstream. Humans have a much more severe reaction than do mice.

One of the main ways that endotoxins enter the animal’s bloodstream is when intestinal integrity is compromised. There can be several reasons for this. The gut can be compromised as the result of diseases such as necrotic enteritis or coccidiosis in chickens or ileitis in pigs. Other factors such as heat stress can also increase the permeability of the intestine and allow increased amounts of endotoxin to enter the bloodstream. The toxicity seen from endotoxins is associated with responses of the animal’s immune system to the LPS in the animal’s bloodstream, once the endotoxins enter the bloodstream they can cause fever, inflammation, and decreased growth. With high levels of endotoxins entering the bloodstream, shock, organ failure, and death can occur.  However, if the endotoxin could be bound and contained in the intestine, it would not have the ability to cause these reactions. 

Calibrin®-Z (available in select international markets), a proprietary thermally processed calcium montmorillonite, has been shown to effectively bind a broad range of bacterial (e.g., alpha toxin and NetB from Clostridium perfringens, E. coli Shiga–like toxins, etc.) and fungal (e.g., zearalenone, fumonisin, aflatoxin, ochratoxin, T-2 toxin, ergovaline, and ergotamine) toxins. Because it binds these toxins, it prevents them from damaging the intestine or from being absorbed through the intestine and entering the body to cause damage to internal organs (e.g., liver, kidneys) hence reducing animal performance. Because of this ability of Calibrin-Z, it was thought that it might also bind lipopolysaccharides. Therefore, a laboratory test was conducted to determine if Calibrin-Z could bind the lipopolysaccharide from E. coli. 

This was done using an in vitro test called the Limulus Amebocyte Lysate assay, which uses the fact that the blood cells (amebocytes) of the horseshoe crab (Limulus) secrete an enzyme (lysate) that clumps when mixed with endotoxins. In preparation for this test LPS solutions with concentrations of 5, 10, 50, or 500 milligrams of LPS per kilogram were made. When these solutions were then treated with Calibrin-Z it resulted in different ratios of Calibrin-Z to LPS, ranging from 20:1 to 10,000:1. The Calibrin-Z was removed using a 0.2 micrometer filter and the remaining solutions were tested using the Limulus Amebocyte Lysate assay to determine the amount of lipopolysaccharide that remained in the solution (Figure 1).

 

These results show that the processed calcium montmorillonite bound the E. coli LPS. The percentage of toxin that was bound increased as the amount of Calibrin-Z to LPS toxin increased. When the binder to toxin ratio was 10,000: 1 Calibrin-Z removed 95% of the LPS from the solution. Calibrin-Z effectively bound lipopolysaccharides, the deadly toxins released when gram-negative bacteria die. Based on this information, feeding Calibrin-Z could help protect poultry and livestock performance from toxic LPS.  To learn more about Calibrin-Z or details about this and other studies, visit amlan.com or contact your local Amlan sales representative.

The Role of Quorum Sensing in Necrotic Enteritis Development

July 11, 2023
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The fast onset of necrotic enteritis and the devastating production losses it inflicts make it one of the most challenging diseases for the poultry industry, particularly for antibiotic-free producers. Clostridium perfringens, the cause of necrotic enteritis, possesses a number of virulence factors that allow it to mount a fast, efficient attack on the host including potent toxins and cell-to-cell communication (quorum sensing). However, natural mineral-based products that can disrupt quorum sensing and reduce the virulence of C. perfringens are available.

The Pathogenesis of Necrotic Enteritis

C. perfringens is an anerobic, spore-forming pathogen found in the normal microbiota of poultry, as well as the poultry house. Necrotic enteritis occurs when predisposing conditions, such as a change in diet, immune status or intestinal pathophysiology, promote an overgrowth of C. perfringens. Coccidiosis can also increase the incidence of necrotic enteritis, as the damage that Eimeria spp. cause to intestinal epithelial cells promotes the invasion of C. perfringens (as well as other pathogens).

C. perfringens’ Rapid Infection Rate

C. perfringens is one of the fastest growing bacterial pathogens. Under optimal conditions it can replicate every 8 to 10 minutes — outgrowing other resident bacteria to achieve intestinal colonization.1 As well as a rapid growth rate, C. perfringens infection involves multiple steps, which likely occur simultaneously, including colonization, replication, nutrient procurement, evasion of host immune defenses, host tissue damage and transmission.2

Exotoxin Roles in Necrotic Enteritis Development

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, while NetB toxin forms pores in cell membranes that allow electrolytes to rupture cells, causing cell death and necrotic lesions in the small intestinal mucosa.3 These two toxins are known to have a role in necrotic enteritis development.

C. perfringens Growth Depends on Host Nutrients

C. perfringens relies on nutrients from the host to live and multiply — a process which results in the destruction of host tissues (formation of necrotic lesions). C. perfringens lacks enzymes needed for amino acid biosynthesis and subsequent protein synthesis, so enzymes and toxins are released to degrade structural proteins from the host.4 The host amino acids and/or peptides are then taken in by C. perfringens for use in its own protein synthesis. 4

To produce energy, C. perfringens degrades large sugar compounds from the host and ferments them, producing gas that enhances the anerobic environment.4 C. perfringens also produces hyaluronidases that increase connective tissue permeability and help C. perfringens spread into deeper tissues.4

Quorum Sensing Controls Exotoxin and Enzyme Production

C. perfringens uses quorum sensing (cell-to-cell communication) to coordinate exotoxin and enzyme production to occur when its population reaches a density that supports the most efficient use of its metabolic resources.5 For example, to determine the optimum time to start producing NetB, the accessory gene regulator-like (Agr-like) quorum-sensing system sends out signals that are recognized by the VirR/VirS two-component regulatory system.5 Once the VirR/VirS system detects that the C. perfringens population has reached the threshold density, it switches on the expression of NetB and other virulence and related metabolism genes.2

Quorum Quenching Reduces Pathogen Virulence

Quorum quenching is an approach that can disrupt the quorum-sensing system of pathogenic bacteria, preventing cell-to-cell communication and the expression of quorum-sensing-controlled genes that produce toxins and other virulence factors. Additionally, quorum-quenching products should reduce the chance of antibiotic resistance, since they are modifying bacteria behavior rather than killing them.

Natural Quorum-Sensing Control

One product that has displayed quorum-quenching properties is the mineral-based feed additive Calibrin®-Z (available in select international markets). This all-natural single-ingredient product binds bacterial pathogens and the toxins they produce, as well as multiple mycotoxins, to help protect the intestinal barrier against enteric disease. Other natural mineral-based products can also help manage necrotic enteritis; read this article to learn more.

An in vitro study found that Calibrin-Z separated out quorum-sensing molecules by adsorption or catalytically broke them down into small fragments. By reducing the concentration of quorum-sensing biochemicals, Calibrin-Z can potentially disrupt the ability of pathogenic bacteria (including C. perfringens) to produce toxins, since quorum sensing controls this function. Calibrin-Z was also shown to effectively bind alpha-toxin and NetB toxin, further reducing the virulence of C. perfringens.

 

The global reduction in the use of in-feed antibiotics has compelled producers to rely on other management methods to maintain a healthy intestinal environment in poultry and reduce the risk of necrotic enteritis. The use of best-practice management strategies and inclusion of mineral-based feed additives that reduce the virulence of C. perfringens can assist in promoting intestinal health and maximizing production efficiency. To learn more about necrotic enteritis and natural mineral-based methods to control it, contact your local Amlan representative.

 

References

  1. Kiu R, Hall LJ. An update on the human and animal enteric pathogen Clostridium perfringens. Emerg Microbes Infect. 2018;7:141.
  2. Prescott JF, Parreira VR, Mehdizadeh Gohari I, Lepp D, Gong J. The pathogenesis of necrotic enteritis in chickens: what we know and what we need to know: a review. Avian Pathol. 2016;45:288–94.
  3. 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.
  4. Shimizu T, Ohtani K, Hirakawa H, Ohshima K, Yamashita A, Shiba T, Ogasawara N, Hattori M, Kuhara S, Hayashi H. Complete genome sequence of Clostridium perfringens, an anaerobic flesh-eater. Proc Natl Acad Sci U S A. 2002;99:996–1001.
  5. Yu Q, Lepp D, Mehdizadeh Gohari I, Wu T, Zhou H, Yin X, Yu H, Prescott JF, Nie SP, Xie MY, Gong J. The Agr-Like Quorum Sensing System Is Required for Pathogenesis of Necrotic Enteritis Caused by Clostridium perfringens in Poultry. Infect Immun. 2017;85:e00975-16.

 

Can dietary inclusion of montmorillonite clay help mitigate the farmed shrimp AHPND pandemic?

June 30, 2023
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Acute Hepatopancreatic Necrosis Disease (AHPND), a prevalent bacterial disease, causes severe economic losses amongst shrimp producers. The shrimp farming industry is currently facing pandemic levels of AHPND. Producers worldwide are at a critical stage where antibiotic-free alternatives are needed. A research trial conducted in Malaysia by Dr. Wing-Keong Ng suggests including a natural-based #montmorillonite clay (Calibrin®-Z) in functional feed improves the growth and health of shrimp. Read the detailed findings published in Global Seafood Alliance.

Calibrin®-A Safeguards Ducklings and Broiler Chicks Against Aflatoxin

June 23, 2023
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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.

 

References

  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.

 

Advanced Technologies to Mitigate Disease in Poultry Production

May 8, 2023
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As more poultry producers incorporate antibiotic-free (ABF) practices into their operations, the need for innovative disease mitigation technologies has also increased. Amlan International is meeting this demand by developing next-generation products, using mineral-based technology, that optimize bird intestinal health and add value for the producer.

In this post we examine four technological advances that could help drive progress in managing disease, bacteria and toxins in flocks while meeting consumer demands for high-quality animal protein. We discuss the advanced mineral technology that is the foundation of Amlan’s natural feed additives as well as a unique diagnostic service offered by Amlan. We also highlight technology on the market from other (unaffiliated) companies that our team thought might be useful for ABF producers to consider when looking for new ways to manage disease in their flock.

  1. Optimize intestinal health with advanced mineral technology

In a crowded feed additive market, advanced mineral technology is the Amlan difference. As the animal health business of Oil-Dri® Corporation of America, Amlan’s scientifically proven products are backed by

Oil-Dri’s 80 years of experience in mineral science. The single-source raw material that is the core of Amlan’s products is calcium montmorillonite with high-capacity opal-CT lepispheres. Proprietary thermal processing — that does not use harmful chemicals — is tailored for each product to create unique functionalities.

Calibrin®-Z (available in select international markets) is a broad-spectrum biotoxin binder that protects poultry and livestock from biotoxins that reduce performance and cause morbidity or mortality. All-natural Calibrin-Z binds polar (e.g., aflatoxin) and nonpolar (e.g., zearalenone) mycotoxins, as well as bacterial toxins like those produced by C. perfringens, E. coli and C. difficile. Binding these biotoxins enhances intestinal health to improve feed efficiency and overall animal performance.

  1. Minimize production loss with timely mycotoxin diagnostic results

When determining whether feed is contaminated with mycotoxins, a fast and accurate analysis is critical for preventing the negative health and production effects mycotoxins can cause. Amlan has partnered with Envirologix, the world’s most trusted name for quantitative testing kits at the point of grain delivery, to bring producers . This program provides on-site quantitative mycotoxin detection results within 10 minutes. The test identifies mycotoxin contamination levels in the feed, and Amlan’s dose calculator can be used to determine the optimal dose of to apply in the animal’s ration to mitigate the toxin threat.

  1. Automation technology increases animal welfare and detects disease

Robot automation in poultry barns is a growing segment of the poultry industry. Robotic technology uses sensors and 2D and 3D cameras to perform repetitive and time-consuming tasks such as disinfecting barns, picking up floor eggs, vaccinating and managing litter.

Increasingly important in modern poultry operations, robotics can also help mitigate production risks and ensure animal welfare goals are met. Thermal imaging can be used to detect animal body temperatures, early disease indicators, record bird activity/welfare and report mortalities while performing their assigned tasks. Monitoring technology can also help producers observe environmental metrics in facilities such as temperature, humidity and carbon dioxide and ammonia levels. A recent development in this area is using sensors to determine when a poultry house smell is no longer “normal.” This technology could help identify a biosecurity or disease risk to a flock.

  1. Blockchain technology facilitates food traceability

Modern consumers are increasingly demanding to know more about how and where their food is raised. Blockchain technology, often in combination with other digital tech, helps track and trace animals and food products throughout the supply chain by linking small packages of data to provide a digital audit of meat production and food safety. At the farm level, these data points could include point of origin, veterinary and feed source records and transportation dates.

As poultry move through the food production system, blockchain technology records significant data points to ensure food supplies meet the traceability requirements that consumers demand. In the processing sector, for example, blockchain can link production information with the final product’s labeling and packaging information to ensure compliance with brand commitments. Blockchain offers incredible promise for connecting poultry growers with new market opportunities.

These emerging trends are important to monitor so producers can determine how best their operations can evolve and remain competitive. Investing in the right technologies will be the key to moving poultry producers to greater production, improved efficiencies, enhanced market opportunities and increased profitability. At Amlan, we are developing and bringing to market advanced feed additive technologies that optimize the intestinal health and production efficiency of producers’ flocks while preserving the economic viability of poultry production. To learn more, click .

Key Necrotic Enteritis Virulence Factors: Alpha-Toxin and NetB toxin

May 8, 2023
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Necrotic enteritis is a common — and costly — infectious disease of poultry caused by the gram-positive bacteria Clostridium perfringens. The virulence of C. perfringens is mostly related to its ability to produce potent toxins. Two exotoxins in particular — alpha-toxin and necrotic enteritis B-like toxin (NetB) — play critical roles in necrotic enteritis development in poultry.

Necrotic Enteritis Origins

The anaerobic, spore-forming C. perfringens is found throughout the poultry environment and is also part of the normal bird microbiota. Necrotic enteritis occurs when there is an overgrowth of C. perfringens caused by predisposing conditions, such as diet changes, immune status or intestinal pathophysiology. High mortality rates are characteristic of the clinical form of necrotic enteritis, whereas subclinical disease causes reduced weight gain and higher feed conversion ratio (FCR). The increased FCR is due to reduced nutrient digestibility and adsorption, and the subsequent compensatory feed intake.

The Role of Exotoxins in Necrotic Enteritis

C. perfringens produces multiple exotoxins, including alpha-toxin and NetB toxin. All C. perfringens types produce alpha-toxin which is cytotoxic to endothelial cells, red blood cells, white blood cells and platelets.1 Alpha-toxin was thought to be the major toxin responsible for necrotic enteritis clinical signs; however, researchers identified an alpha-toxin-negative mutant strain that was able to induce necrotic enteritis.2 This led to the discovery that NetB was a key virulence factor of necrotic enteritis in poultry.

Quorum sensing regulates NetB production, which is initiated when C. perfringens reaches a concentration of 109 CFU/g or higher and the bird has dysbiosis or impaired intestinal function.3 In healthy birds, the intestinal epithelium and mucus layer protect internal tissues by forming a selective barrier against the contents of the intestinal lumen (the external environment). The natural barrier allows nutrients through the intestinal wall but not pathogens and their toxins. A breakdown of this defensive barrier occurs when there is an overgrowth of C. perfringens and toxin production is stimulated. NetB forms pores in cell membranes that allow electrolytes to rupture cells, causing cell death and necrotic lesions in the small intestinal mucosa.1 The damage to the intestinal wall hinders nutrient digestion and absorption and can allow toxins to enter the circulatory system.

Manage Necrotic Enteritis with Natural Mineral-Based Feed Additives

Effective management programs are important for reducing the harmful health and production effects of necrotic enteritis. However, neutralizing the toxins produced by C. perfringens can also help control necrotic enteritis. The natural mineral-based feed additives Varium®, NeutraPath® and Calibrin®-Z (all available in select international markets) can be used alone or in synergistic combination to help maintain a healthy intestinal environment and protect birds from the devastating effects of alpha-toxin and NetB toxin.

Designed specifically for poultry, patented Varium promotes bird efficiency and productivity by protecting the intestinal lining, strengthening the intestinal barrier and stimulating the intestinal immune system. Varium’s synergistic, mineral-based formulation does this by binding pathogenic bacteria and the toxins they produce, energizing intestinal epithelial cells and acting as an immunomodulator to help birds naturally combat disease.

NeutraPath helps support broiler gut health and structural integrity by reducing the intestinal load and colonization of pathogenic bacteria. NeutraPath has bacteriostatic and bactericidal properties and can also neutralize virulence factors, including the toxins produced by pathogenic bacteria and the quorum-sensing molecules used for pathogen communication.

Calibrin®-Z is a single-ingredient, all-natural biotoxin binder that helps protect the intestinal barrier against enteric disease. Calibrin-Z can bind a broad spectrum of pathogens and bacterial and fungal toxins, including C. perfringens and E. coli toxins and aflatoxin, fumonisin and zearalenone.

Natural Mineral-Based Feed Additives Bind C. perfringens-produced Toxins

were shown to effectively bind alpha-toxin and NetB toxin in vitro (Figure 1), and NeutraPath caused a four-fold reduction in unbound alpha-toxin in the cecal contents of C. perfringens-challenged broilers (Figure 2). These alpha-toxin and NetB toxin binding abilities also translated to health and performance improvements during .

Figure 1: Researchers at the United States Department of Agriculture – Agricultural Research Service (USDA-ARS) showed that Varium and Calibrin-Z were able to effectively bind alpha-toxin and NetB toxin in vitro.

Figure 2: A four-fold reduction in unbound alpha-toxin occurred in cecal contents of C. perfringens-challenged broilers fed NeutraPath compared to challenged control birds (128 total birds). Source: Southern Poultry Research, Inc., Athens, GA.

The C. perfringens toxins associated with the development of necrotic enteritis — alpha-toxin and NetB toxin — can have devastating effects on the health and production performance of poultry. However, the natural mineral-based feed additives Varium, NeutraPath and Calibrin-Z can help neutralize these toxins, protect the intestinal environment and improve production efficiency. To learn more about these intestinal health solutions, contact your local Amlan representative.

References

  1. 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.
  2. Keyburn AL, Sheedy S, Ford M, Williamson M, Awad M, Rood J, Moore RJ. Alpha-toxin of Clostridium perfringens is not an essential virulence factor in necrotic enteritis in chickens. Infect Immun. 2006;74:6496-6500.
  3. Mora ZV, Macías-Rodríguez ME, Arratia-Quijada J, Gonzalez-Torres YS, Nuño K, Villarruel-López A. Clostridium perfringensas Foodborne Pathogen in Broiler Production: Pathophysiology and Potential Strategies for Controlling Necrotic Enteritis. Animals (Basel). 2020;10:1718.

Natural Mineral-Based Solutions for Managing Necrotic Enteritis in Poultry

March 29, 2023
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Necrotic enteritis causes economic loss for poultry producers around the world due to bird mortality and production inefficiency. The increasing restrictions on antibiotic growth promoter use have complicated the issue of necrotic enteritis control for antibiotic-free (ABF) producers. However, management programs and mineral-based feed additives can provide natural control methods to reduce the incidence of necrotic enteritis in flocks.

What Is Necrotic Enteritis?

Necrotic enteritis is caused by Clostridium perfringens, a spore-forming anerobic bacteria found throughout the poultry house environment and the normal microbiota of the bird. Disease in birds occurs when there are predisposing conditions, such as diet changes, immune status or intestinal pathophysiology, that cause an overgrowth of C. perfringens. The exotoxins alpha-toxin and necrotic enteritis B-like toxin (NetB) are produced by C. perfringens and have a role in necrotic enteritis development.

Necrotic enteritis occurs due to a breakdown of the defensive intestinal barrier (epithelium and mucus layer). The damage to the intestinal wall impairs nutrient digestion and absorption and can allow toxins to enter the circulatory system. Coccidiosis, caused by Eimeria spp., can increase the incidence of necrotic enteritis as Eimeria also damage intestinal cells. Mycotoxicosis from feed contaminated with fungal toxins can also increase the negative effects of necrotic enteritis. High mortality is a characteristic of clinical necrotic enteritis, while decreased weight gain and reduced feed efficiency are associated with subclinical disease.

 

Management Programs to Control Necrotic Enteritis

With the global push toward ABF production, maintaining a healthy intestinal environment to reduce the risk of necrotic enteritis now requires effective management strategies. Reduced ventilation, increased litter moisture and poor husbandry can increase the incidence of necrotic enteritis. However, natural feed additives that strengthen and protect the intestinal environment can also help reduce the risk of necrotic enteritis.

Natural Mineral-Based Feed Additives

Along with management best practices, the natural mineral-based feed additives Calibrin®-Z, Varium®, and NeutraPath® (all available in select international markets) can help maintain a healthy intestinal environment and reduce the incidence of necrotic enteritis. Third-party research has shown these products improve the health and performance of necrotic enteritis-challenged broilers.

Calibrin-Z for Broad-Spectrum Biotoxin Defense

Single-ingredient Calibrin-Z protects the intestinal barrier against damage from a broad spectrum of pathogens and bacterial and fungal toxins, including C. perfringens and E. coli toxins and aflatoxin, fumonisin and zearalenone. During manufacture, mineral-based Calibrin-Z undergoes proprietary thermal processing that permits multiple binding mechanisms, including hydrophobic interactions, chelation, hydrogen bonding and electrostatic attraction. The result is enhanced pathogen and toxin adsorption without significant binding of important nutrients.

In a study published in the Journal of Applied Poultry Research, Calibrin-Z reduced the effects of necrotic enteritis challenge, including returning gain during days 10 to 24 to levels equal to non-challenged birds. When birds were challenged with necrotic enteritis plus dietary aflatoxin (1 mg/kg), it significantly increased the negative effects of necrotic enteritis. However, Calibrin-Z improved feed efficiency to levels equivalent to birds challenged with necrotic enteritis only. Virginiamycin was not able to do this, demonstrating the benefits of feeding Calibrin-Z, which can bind mycotoxins and bacterial pathogens and their toxins.

Varium Promotes Poultry Efficiency and Productivity

Varium meets the poultry industry’s need to reduce the use of in-feed antibiotic growth promoters while maintaining bird health and performance and providing added-value for producers. Varium’s patented formulation includes Amlan’s mineral technology, yeast and a functional amino acid that work synergistically to reduce pathogenic challenges, strengthen the intestinal barrier and prepare the immune system to naturally defend against disease. With its multiple modes of action, Varium can replace the need for multiple feed additives by providing the same benefits in one product, reducing feed costs and simplifying diet formulations.

Multiple broiler studies have proven that Varium can be as effective as antibiotics in maintaining performance during necrotic enteritis challenge. In a study where necrotic enteritis-challenged broilers were fed for 28 days, feed conversion ratio (FCR), weight gain and mortality were similar between Varium and the antibiotic growth promoter bacitracin methylene disalicylate (BMD; Figure 1). Similar results were observed when Varium was compared to virginiamycin in another necrotic enteritis challenge study (Figure 2). These results show the value Varium can add to ABF systems by protecting the intestinal environment and promoting bird efficiency.

Figure 1: Weight gain and feed conversion ratio (FCR) of broilers fed BMD, Varium or non-supplemented for 28 days and undergoing necrotic enteritis challenge.

Figure 2: Weight gain and feed conversion ratio (FCR) of broilers fed virginiamycin, Varium or non-supplemented for 28 days and undergoing necrotic enteritis challenge.

NeutraPath for Natural Pathogen Control

Another natural option for managing necrotic enteritis is NeutraPath — a proprietary and co-active blend of essential oils, fatty acids and Amlan’s mineral technology. The synergistic formula of NeutraPath is designed to have multiple modes of action, resulting in enhanced intestinal health, improved feed efficiency and reduced mortality in necrotic enteritis-challenged birds. In environments with high bacterial challenge, NeutraPath can reduce the intestinal load and colonization of pathogenic bacteria and help maintain broiler gut health and structural integrity.

NeutraPath has bacteriostatic and bactericidal properties that target cell function and cell wall integrity of pathogenic bacteria. It also neutralizes virulence factors, including the toxins produced by pathogenic bacteria and the quorum-sensing molecules used for pathogen communication.

The on-farm benefits of pathogen control by NeutraPath have been demonstrated in multiple broiler trials. For example, in a 28-day trial that challenged broilers with C. perfringens and Eimeria, NeutraPath reduced mortality and lesion scores to levels similar to birds administered BMD (Figure 3). Additionally, broilers in the NeutraPath group had greater weight gain and a better FCR for the overall 28-day period than necrotic-enteritis challenged control birds, and NeutraPath-fed birds did not differ statistically from BMD-treated broilers (Figure 4). This study shows the performance benefits achieved with NeutraPath by improving intestinal health and reducing the intestinal pathogen load.

Figure 3: NeutraPath reduced mortality and lesions scores by half compared to the necrotic enteritis-challenged control. abcMeans with different superscripts are significantly different (P < 0.05).

Figure 4: NeutraPath improved weight gain and feed efficiency compared to the necrotic enteritis-challenged control. abMeans with different superscripts are significantly different (P < 0.05).

 

An increasing number of poultry producers are moving toward the global objective of reduced in-feed use of antibiotics. Consequently, maintaining a healthy intestinal environment now relies more heavily on other management methods to reduce the risk of necrotic enteritis. 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 and the natural mineral-based feed additives that can help manage it, contact your local Amlan representative.

 

 

Managing Post-Weaning Diarrhea in Pigs with Natural Feed Additives

March 13, 2023
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The performance and health setbacks, like post-weaning diarrhea, that pigs face during weaning (the fallback effect) can affect growth across the pig’s lifetime and cause significant economic loss for producers. Bacterial toxins are common contributors to post-weaning diarrhea, but natural solutions are available that can target these toxins and strengthen the intestinal barrier, helping to reduce the incidence of diarrhea and keep weaned piglet performance on track.

E. coli Toxins: A Cause of Post-Weaning Diarrhea

Escherichia coli (E. coli) produces potent toxins that can cause enteric disease in recently weaned pigs. Enterotoxigenic Escherichia coli (ETEC) is a common cause of post-weaning diarrhea. It produces heat-labile enterotoxin (LT) that alters the electrolyte and water balance in intestinal cells, leading to diarrhea.1 A survey of U.S. ETEC isolates linked to porcine post-weaning diarrhea showed that 57.7% had the heat-labile toxin gene.2

Additionally, when E. coli die, they release endotoxin (lipopolysaccharides, LPS) from the cell wall. Endotoxin can damage intestinal cells leading to weakness in the intestinal barrier that can cause “leaky gut” syndrome, causing diarrhea and allowing endotoxin to enter the bloodstream where it can cause shock or even death.

Calibrin®-Z Is a Natural Binder of Bacterial Toxins

Mineral-based Calibrin®-Z (available in select international markets) helps protect the intestinal environment of swine from a broad spectrum of pathogens and bacterial toxins (including E. coli toxins), as well as fungal toxins like aflatoxin, ochratoxin, fumonisin and zearalenone. Single-ingredient Calibrin-Z undergoes specifically tailored proprietary thermal processing during manufacture that optimizes its binding ability. The result is enhanced bacterial and fungal toxin adsorption without significant binding of important nutrients.

Multiple in vitro studies by third-party laboratories have proven the bacterial toxin binding capabilities of Calibrin-Z. The toxins bound by Calibrin-Z in vitro include E. coli heat-labile enterotoxin and E. coli endotoxin (LPS), as well as difficile (formerly known as Clostridium difficile) toxins, toxin A and toxin B, a cause of enteric disease in neonatal piglets (Figure 1).

Figure 1: Calibrin-Z bound multiple E. coli and C. difficile toxins in vitro.

NeoPrime® for Naturally Healthier Weaning

Traditionally, antibiotics were included in nursery pig diet formulations to improve performance and reduce the risk of disease during weaning. However, concern over antimicrobial resistant pathogens and consumer preferences have reduced the prophylactic use of antibiotics in swine diets. Similarly, high levels of zinc oxide (2,000 to 3,000 ppm) were often added to weaned piglet rations to decrease the incidence of diarrhea and improve weight gain. But this practice has raised concerns of overloading zinc in soil and water when manure is used as fertilizer.

A natural alternative to antibiotic and zinc oxide use is the mineral-based feed additive NeoPrime® (available in select international markets). To counteract the performance fallback effects of weaning, patented NeoPrime positively modifies the intestinal environment, energizes enterocytes for better nutrient absorption and primes the intestinal immune system. The ingredients in NeoPrime — Amlan’s mineral technology plus yeast and a functional amino acid — work synergistically to reduce pathogenic challenges, strengthen the intestinal barrier and safely stimulate the intestinal immune system.

A study conducted by Chi et. al (2018) looked at the performance benefits of adding NeoPrime to a diet that included antibiotics and zinc oxide, as well as the effects when antibiotics, zinc oxide or their combination were removed from the diet.3 The researchers also monitored E. coli endotoxin concentration in blood plasma.

Weaned pigs (90 male, 90 female) 28 days of age (6.8 kg) were randomly assigned to five treatments with six replications. Pigs were fed one of five treatments (Table 1) for 35 days, with feed consumption and body weight recorded on days 14 and 35. The antibiotics used were chlortetracycline (75 g/MT) and olaquindox (100 g/MT), and the zinc oxide was included at a rate of 3 kg/MT. Blood was drawn from one pig per pen on days 14 and 35.

Table 1: Piglets were fed one of five treatments with or without NeoPrime, antibiotics and zinc oxide.

 

On day 14, pigs fed NeoPrime maintained body weight and feed conversion, even when antibiotics, zinc oxide or their combination were removed from the diet (Table 2). By day 35, pigs fed NeoPrime tended to increase body weight instead of only maintaining performance and had numerically improved feed conversion compared to the control (Table 2). Plasma endotoxin was generally lower when NeoPrime was added to the diet (Figure 2), indicating that NeoPrime bound and removed LPS from the intestine. The research suggests that NeoPrime is an effective natural alternative to prophylactic antibiotic use and zinc oxide supplementation.

Table 2: NeoPrime maintained performance even when antibiotics, zinc oxide or their combination were removed from the diet.

 

Figure 2: NeoPrime decreased plasma endotoxin compared to the control.

 

NeutraPath for High-Challenge Environments

In high-challenge environments, swine can be fed NeutraPath® (available in select international markets) which is a proprietary and co-active blend of essential oils, fatty acids and Amlan’s mineral technology. NeutraPath has multiple modes of action, including , decreasing pathogenic bacteria with its bacteriostatic and bactericidal properties and improving intestinal structural integrity. By reducing pathogenic bacterial intestinal colonization, NeutraPath can improve intestinal health and help reduce the incidence of post-weaning diarrhea.

The health and performance benefits of NeutraPath in ETEC-challenged pigs were shown in a study by He et al. (2022).4 Pre-weaned piglets (18 male, 18 female) were enrolled in the study after fecal samples were confirmed to be absent from β-hemolytic E. coli and genotyping verified susceptibility to F18 ETEC. Weaned piglets were fed a control diet, or that diet supplemented with 0.25% or 0.5% NeutraPath for 28 days. After a 7-day adaptation period, pigs were orally inoculated with F18 ETEC. Fecal consistency was scored twice daily after inoculation, and fecal and gut microbiome samples were taken at multiple time points during the challenge period.

Overall, NeutraPath reduced the incidence and severity of diarrhea. The NeutraPath groups had lower (P < 0.05) frequency of diarrhea score ≧ 3 than the control group (Figure 3) and the NeutraPath 0.25% group had lower frequency of diarrhea score ≧ 4 than the control (Figure 3).

Figure 3: NeutraPath reduced the overall incidence and severity of diarrhea.

Both NeutraPath dose rates improved (P = 0.048) gain:feed from day 14 to 21 post-infection (the recovery period) and improved (P < 0.05) growth across the whole feed period regardless of dose. The overall growth rates were 258% (NeutraPath 0.25%), 261% (NeutraPath 0.5%) and 229% (control) higher than the weight at the day of the challenge.

The study also showed that NeutraPath may help maintain desirable bacteria in the intestine during the ETEC infection period (Figure 4). On day 7 post-infection, both NeutraPath groups had a higher relative abundance of Lactobacillaceae and reduced relative abundance of Bacteroidaceae and Enterobacteriaceae in fecal samples. The NeutraPath 0.5% group also had increased relative abundance of Lachnospiraceae in feces on day 7 post-infection and enhanced relative abundance of Clostridiaceae1 and reduced Enterobacteriaceae in the ileal mucosa.

 

Figure 4: NeutraPath helped maintain desirable bacteria in the intestine during the ETEC infection period.

 

Changes to the fecal microbiome and ileal mucosa microbiota composition indicated that NeutraPath may be able to maintain a more favorable intestinal environment during ETEC infection. NeutraPath also reduced the incidence and severity of diarrhea and had better performance during the recovery period in this study, suggesting NeutraPath is an effective natural method for managing post-weaning diarrhea.

 

Amlan offers swine producers a range of natural options to help manage post-weaning diarrhea in piglets. Our products can reduce the pathogen load in the intestinal environment and strengthen the intestine, keeping weaned piglets healthier and more productive. For more information on our range of natural solutions for post-weaning diarrhea, contact your local Amlan representative.

 

References

  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;9:292. https://doi.org/10.3389/fcimb.2019.00292
  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. Vet Microbiol. 2007;123:145–152. https://doi.org/10.1016/j.vetmic.2007.02.018
  3. Chi F, Johnston S, Tang X, Chen W, Wang B, Tang S. PSIV-28 Effects of replacing zinc oxide and antibiotics with NeoPrime® on growth performance and plasma and fecal endotoxin concentration in nursery pigs. J Anim Sci. 2018;96(Suppl 3):321. https://doi.org/10.1093/jas/sky404.706
  4. He Y, Jinno C, Li C, Johnston SL, Xue H, Liu Y, Ji P. Effects of a blend of essential oils, medium-chain fatty acids, and a toxin-adsorbing mineral on diarrhea and gut microbiome of weanling pigs experimentally infected with a pathogenic Escherichia coli. J Anim Sci. 2022;100(1):skab365. https://doi.org/10.1093/jas/skab365

A Natural Solution to Mitigating Acute Hepatopancreatic Necrosis Disease (AHPND) in Shrimp

February 22, 2023
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Acute hepatopancreatic necrosis disease (AHPND), originally called “early mortality syndrome,” costs the shrimp industry billions of dollars each year. Outbreaks of the disease have occurred in multiple countries around the world, leading to shrimp mortality and loss of income for many commercial farms. Here we discuss the etiology of AHPND and a natural solution for mitigating its effects.

The Bacteria Behind AHPND

The causative agent of AHPND is Vibrio parahaemolyticus, an orally transmitted bacteria that colonizes the shrimp gastrointestinal tract. V. parahaemolyticus produces a potent binary toxin that enters the hepatopancreas and causes dysfunction and destruction of this vital organ, often leading to mortality. Affected shrimp present with an abnormal hepatopancreas that is often reduced in size and shows whitening and hardening. However, very high mortality within a short period of time may be the first indication that a pond has been infected.

A Toxic Cause of Hepatopancreatic Cell Death

The specific strain of V. parahaemolyticus that causes AHPND was first identified by Dr. Lightner’s group at the University of Arizona, who also reported that the pathology was caused by a toxin secreted by V. parahaemolyticus.1 Lee et al. (2015) showed that the strains of V. parahaemolyticus that cause AHPND contain a plasmid (pVA1) that encodes the binary toxin PirABvp (Photorhabdus insect-related toxins PirAvp and PirBvp) and that this toxin leads to the death of shrimp hepatopancreatic cells.2

Natural Protection from AHPND

Protecting shrimp from the damaging effects of AHPND requires consideration of multiple factors, including biosecurity measures and other production practices that reduce the risk of disease. However, a natural mineral-based solution, Calibrin®-Z (available in select international markets), is also available to help protect shrimp from AHPND and the devastating production losses it causes.

Calibrin-Z is a broad-spectrum biotoxin binder manufactured from a single ingredient — Amlan’s unique mineral technology. Calibrin-Z is optimized via proprietary thermal processing and binds mycotoxins and certain endo and exotoxins produced by pathogenic bacteria, helping to defend the intestinal tract from enteric disease.

When bacterial toxins (and mycotoxins) flow through the gastrointestinal tract, they are bound by Calibrin-Z and thus prevented from entering the hepatopancreas. The bound toxin is removed through normal digestive function (Figure 1). To provide optimal protection against AHPND, Calibrin-Z should be fed consistently from the beginning to the end of the growing cycle.

Figure 1: Calibrin-Z helps prevent PirA and PirB toxins from entering the hepatopancreas by binding the toxins. The bound toxin is removed through normal digestive function.

Research Proves Protective Effect of Calibrin-Z

A number of studies have shown the protective benefits of adding Calibrin-Z to shrimp diets. Here we present a commercial farm case study as well as two university-based studies that demonstrate the health, performance and economic benefits of Calibrin-Z.

Commercial Case Study Shows Strong ROI

In this study at a commercial farm in Mexico, shrimp were fed either probiotics and charcoal (control group, five ponds) or Calibrin-Z (5 kg/MT feed, four ponds) from post-larvae to market weight (approximately 90 days). Each pond (5 ha) was stocked with 1 million shrimp. An AHPND outbreak did not occur during this trial.

Shrimp fed Calibrin-Z had greater average weekly weight gain (P < 0.1), improved feed conversion at harvest (P < 0.05) and reached harvest time (14 g weight) six days earlier (Figure 2).

Figure 2: Calibrin-Z improved shrimp performance compared to shrimp fed probiotics and charcoal.

Calibrin-Z Provides Hepatopancreatic Protection and Increases Survival Rates

Another study was conducted at Instituto Tecnológico de Sonora, Laboratorio de Analisis en Sanidad Acuícola (Aquatic Health Laboratory) in Obregon, Sonora, Mexico. White shrimp (Penaeus vannamei, 30 shrimp per treatment, three replications per treatment) were fed one of four treatments: negative control (no challenge), positive control (V. parahaemolyticus challenged), Calibrin-Z at 0.25% (challenged) or Calibrin-Z at 0.5% (challenged). The shrimp were fed twice a day for 20 days (fed for 15 days then challenged for 5 days). The positive control and Calibrin-Z groups were challenged by immersion with bacterial broth (TSB) containing V. parahaemolyticus (AHPND, strain 6V) via waterborne contact infection.

Calibrin-Z fed at 0.5% doubled the survival rate after experimental V. parahaemolyticus infection compared to the positive control (63.3% vs. 30%, Figure 3) and significantly improved cumulative mortality (Figure 4). A protective effect in the hepatopancreas was also observed in shrimp fed 0.5% Calibrin-Z compared to the positive control. Severity of AHPND histopathology (G0 = an absence of damage and G4 = severe tissue lesions) was 0 for the negative control, 4 for the positive control and 2 for Calibrin-Z fed at 0.5%. Calibrin-Z fed at 0.5% clearly shows significant improvement in shrimp survival rates.

Figure 3: Calibrin-Z fed at 0.5% doubled the survival rate of shrimp challenged with V. parahaemolyticus infection compared to the positive control (P < 0.05).

 

Figure 4: Cumulative mortality was improved by feeding Calibrin-Z to shrimp challenged with V. parahaemolyticus compared to the challenged control.

 

Enhanced Microbiota Diversity and Improved Survivability with Calibrin-Z

A study conducted at Universiti Sains Malaysia, Penang, Malaysia, looked at the benefits of Calibrin-Z on post-larval growth, gut health and disease resistance to AHPND. The researchers used triplicate groups of specific pathogen-free shrimp (Penaeus vannamei, 20 per replicate) and compared the performance and health of a negative control, positive control and Calibrin-Z fed at 0.25% or 0.5%. All groups were fed for 14 days and then a 7-day emersion challenge was conducted with all groups, except the negative control, using an AHPND-causing strain of V. parahaemolyticus.

Calibrin-Z at 0.25 or 0.5% did not affect growth, feed efficiency or survival during the 14-day unchallenged period. When challenged, shrimp fed Calibrin-Z at 0.25% and 0.5% had significantly higher survival rates (83.3 and 93.8%, respectively) compared to the positive control group (39.6%) and were not significantly different to the negative control (96.7%).

Presumptive Vibrio and total cultivable bacteria counts in the hepatopancreas were lower for shrimp fed Calibrin-Z, and histopathology of the hepatopancreas showed less damage compared to the positive control group. Calibrin-Z also enhanced the stomach microbiota diversity and appeared to modulate the bacterial community, which may have positively affected shrimp survival.

 

These studies demonstrate the benefits Calibrin-Z offers shrimp farmers as a natural solution for mitigating the production and profit-limiting effects of AHPND. The unique biotoxin-binding features of Calibrin-Z help provide protection against bacterial toxins like PirA and PirB as well as mycotoxins in the feed. For more information on how Calibrin-Z can benefit your farm, visit amlan.com or contact your local Amlan representative.

 

 

References

  1. Tran L, Nunan L, Redman RM, Mohney LL, Pantoja CR, Fitzsimmons K, Lightner DV. Determination of the infectious nature of the agent of acute hepatopancreatic necrosis syndrome affecting penaeid shrimp. Dis Aquat Org. 2013;105:45–55. https://doi.org/10.3354/dao02621.
  2. Lee CT, Chen IT, Yang YT, Ko TP, Huang YT, Huang JY, Huang MF, Lin SJ, Chen CY, Lin SS, Lightner DV, Wang HC, Wang AH, Wang HC, Hor LI, Lo CF. The opportunistic marine pathogen Vibrio parahaemolyticus becomes virulent by acquiring a plasmid that expresses a deadly toxin. Proc Natl Acad Sci USA. 2015;112:10798–10803. https://doi.org/10.1073/pnas.1503129112.

 

 

Amlan International Featured in Feed & Additive Magazine

January 19, 2023
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Feed & Additive Magazine covered Amlan’s latest study in their January issue focused on antibiotic reduction strategies and alternative feed additives. The article highlights research backed performance benefits of feeding natural, mineral-based additives with synergistic modes of action when removing antibiotics. As many producers look to offset decreased feed efficiency and increased mortality when removing AGPs, Amlan’s unique mineral technology may be the answer. Click here to learn more.

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