The addition of nucleotides to the diet helps to prevent intestinal dysbiosis. By averting enteritis, nucleotides create the conditions for obtain excellent conversion rates and long-lasting productivity.

Author: Pier Enrico Rossi, DVM

For decades, bacitracin, lincomycin, avoparcin, virginiamycin, tylosin, and avilomycin have enabled the control of necrotic enteritis. Subsequently, regulations aimed at reducing growth-promoting antibiotics, anticoccidials, and ionophores have led to a resurgence of this disease.

The etiological agent of necrotic enteritis is Clostridium perfringens, a Gram-positive, ubiquitous, anaerobic, and spore-forming bacterium. The spores enable Clostridium perfringens to survive in livestock facilities for a long period, facilitating transmission across successive production cycles. In this way it becomes impossible to avoid contact between animals and the bacterium.

Clostridium perfringens strain differ in virulence and their ability to cause disease. The pathogenic strains are able to produce toxins, bacteriocins, and hydrolytic enzymes, enabling them to adhere, form biofilms, and colonize the intestine.

Necrotic enteritis can manifest itself in both clinical and subclinical form. The disease may onset between 2 and 4 weeks of age. The clinical evident phase is short and characterized by depression, ruffled feathers, locomotor ataxia, diarrhoea, and death within a few hours, with a mortality rate that can reach 1% per day.

The subclinical form, difficult to recognize, is the economically most dangerous form; if untreated, it will negatively impact conversion rates and group uniformity. Although it is well-established that pathogenic strains can alter the normal intestinal microflora by replacing commensal (non-pathogenic) clostridia, the only presence of Clostridium perfringens is not sufficient to trigger necrotic enteritis. It is essential an intestinal wall already damaged, as Clostridium perfringens produces alpha-toxin, a phospholipase, a mucolytic enzyme, that allows it to be feed by mucus.

If upstream of Clostridium replication there is an event that triggers increased mucus secretion, the damaged intestine produces T lymphocytes, cytokines, interferon, and interleukins that act on the mucous goblet cells of the intestinal epithelium, activating them and enhancing mucus secretion, thus enabling the proliferation of Clostridium perfringens.

There are many agents that can create damages to the intestinal mucosa: enteric viruses, helminths, mycotoxins, biogenic amines, fat rancidity, and stress. However, the most prevalent pathology is duodenal and jejunal coccidiosis, caused by Eimeria maxima and Eimeria acervuline, which more frequently induce damage to broilers intestines.

Coccidiosis can occur due to increased resistance to coccidiostats or following the use of live vaccines against coccidiosis. These vaccines consist of non-attenuated or minimally attenuated Eimeria species, which must invade the intestinal mucosa and replicate. However, to induce robust immunity, they require three consecutive cycles of infection, risking epithelial damage.

To control coccidiosis and reduce the incidence of necrotic enteritis, a proper rotation of anticoccidial drugs is necessary to preserve their effectiveness. Implementing rotation between drugs and vaccination, based on risk levels and seasonal incidence, along with proper brooder management to promote adequate cycling of oocysts, is crucial to avoid adverse effects and prepare the farm for the occurrence of necrotic enteritis.

The effectiveness of certain polyphenols, combined with plant extracts and minerals, has proven to be effective in reducing the bacterial count in the litter after faecal excretion. This could be a valuable tool in reducing the change of reinfection by Clostridium perfringens.

To prevent alterations that may favour necrotic enteritis, the use of probiotics has become widespread to balance the intestinal microflora. It has been demonstrated that certain strains of lactobacilli—used either alone or in combination with organic acids (formic, propionic, and butyric)—reduce the severity of lesions from haemorrhagic enteritis and the resulting mortality.

Other alternative products, such as MOS or beta-glucans, have demonstrated the ability to reduce the severity of lesions caused by necrotic enteritis due to their anti-inflammatory effects. They promote the secretion of immunoglobulins and the production of short-chain fatty acids (SCFA). It is important remark that Clostridium perfringens, like other enteric pathogens, is a highly adaptable microorganism. Therefore, it is necessary to use these products in a balanced way to prevent the development of resistance phenomena.

A promising new opportunity is represented by the use of nucleotides.

The term nucleotide describes a substance composed of a sugar, a phosphate group, and a nitrogenous base; the bases are purines and pyrimidines, and the sugar is either ribose or deoxyribose. Nucleotides represent the building blocks of RNA and DNA. Additionally, nucleotides are the constituents of molecules such as ATP, NADP, and NADH, responsible for transferring energy produced by the metabolism of carbohydrates, lipids, and proteins.

In a standard condition, nucleotides are not considered essential nutrients, because they can be synthetized by the organism. Nucleotides become essential in case of insufficient synthesis, specifically during growth and in the presence of pathologies. In particular, rapidly turning-over tissues, such as those in the intestinal mucosa, are unable to produce sufficient quantities of nucleotides to meet their needs. It has been demonstrated that nucleotides provided through nutrition act as growth factors on intestinal cells, promoting their differentiation and maturation. This results in increased thickness of the intestinal mucosa, increased villous height and deeper crypts.

Several studies have demonstrated that the supply of nucleotides through nutrition promotes the increase in digestive enzyme activity and stimulates the growth of Lactobacilli and Bifidobacteria, at the expense of Gram-negative bacteria such as Escherichia coli. It is the positive effect of nucleotides on Lactobacilli and Bifidobacteria seems to be due to the increased absorption of trace elements in ionic form by enterobacteria.

In addition, the supply of nucleotides has proven to be very useful in the clinical and functional recovery of liver conditions. Nucleotides promote hepatocyte activity through mitochondrial stimulation aimed at stimulating sirtuin production, accelerating protein synthesis and glucose deposition. This prevents the accumulation of cholesterol and other lipids, reducing the risk of hepatic steatosis. In this way, nucleotides can improve animal productive life extension, especially for laying hens.

While it is true that nucleotides are not considered essential because they can be synthesized from simple substances, it is true, however, that in specific phases, the metabolic synthesis processes may not be able to meet their demand.
For example, during periods of rapid cell turnover, such as the normal growth process of the chick. In the first seven days of life, of which the first three define a significant portion of its subsequent performance, the chick grows by about fourfold.

Although nature provides the chick with nutrients through the residue of the yolk sac, this food reserve is insufficient to face the challenges and stress of the very first days of life.

Having a source of soluble nucleotides derived from yeast extraction allows for administration both in drinking water and in feed, right from the very first hours of life. This enables the chick to reach its maximum growth potential through proper development of the enteric system and its correct microbiota.

The benefits become immediately apparent, with a lower viscosity of feces in the first days of life, resulting in a reduced incidence of anal pasting. The litter will also be drier in the early days, showing a clear reduction in foot pad dermatitis (FPD). Furthermore, the growth will demonstrate group uniformity with an improved conversion rate.

The advantages listed so far are particularly evident in broilers, while in the breeding of long-lived animals such as laying hens, the benefits will stem from improved intestinal well-being, the restoration of liver activity, and the ability of nucleotides, once inside a cell, to generate NAD (nicotinamide adenine dinucleotide), a molecule essential for the activity of more than 500 enzymes, including sirtuins. Sirtuin deficiency is a cause of aging. All of this will result in a better conversion rate, a lower percentage of dirty eggs, a reduction in shell fragility, and the extension of productive activity.

 Introduction

Brazil, indeed a blessed nation with a coastline stretching over 8,500 kilometers and, according to sources, holding between 8% and 13.7% of all the freshwater available on the planet, is rightfully positioned as a promising superpower in the global aquaculture context. Despite its challenges, the national aquaculture sector has seen exponential growth in recent years, particularly in shrimp and fish production, driven by both domestic and international demand.

As Brazil’s aquaculture industry continues to expand, it faces various challenges, including the need to meet this growing demand in an environmentally sustainable and economically viable manner. This brings us to a crucial aspect, the optimization of specially formulated diets to meet the specific requirements of cultured shrimp and fish. The composition and quality of raw materials, ingredients, vitamins, and minerals used in aquaculture feed production require special attention from manufacturers, as their quality and efficiency are closely related to performance.

In times of fishmeal and fish oil scarcity and increasing use of vegetable meals, innovative solutions have been transforming shrimp and fish production: the use of nucleotide-enriched diets. These micro components play a pivotal role in the zootechnical performance of aquatic organisms, the robustness of their immune system, and exhibit strong antioxidant properties.

In this context, at Prosol, we invite everyone to embark on this fascinating journey with us to delve a little deeper into what nucleotides are and the role they play in modern aquaculture. This is the path to a new era of success in shrimp and fish production, driven by science and innovation.

Nucleotides are the building blocks of life. They are organic molecules composed of sugar, a phosphate group, and a nitrogenous base, playing a crucial role in the genetic composition of all living organisms. In essence, nucleotides are the alphabet of the genetic code, forming sequences that make up DNA and RNA. This genetic information serves as the blueprint for the growth, development, reproduction, and functioning of every living organism.

Just for curiosity’s sake, each new cell to be produced requires about ONE BILLION nucleotides to duplicate. In the early stages of aquatic organisms, under conditions of stress and environmental imbalance, during vaccination (in the case of fish), and in cases of tissue damage (such as gills, for example), nucleotide supplementation in feed becomes necessary to ensure that cell multiplication processes meet the specific demands of each challenging situation.

During cell division, DNA is replicated to ensure that each daughter cell inherits the same genetic information as the parent cell. RNA, on the other hand, is essential for protein synthesis, translating DNA information into sequences of amino acids that form proteins, thus playing a vital role in cell function and structure.

Together, DNA and RNA are fundamental to cell division, multiplication, and daily cell operations. Therefore, as essential components of both DNA and RNA, nucleotides play a critical role in improving the overall physiology of cultured organisms, disease prevention and recovery, stress reduction, and promoting better growth rates.

Nucleotides as Health Boosters

Nucleotides act as conductors, orchestrating the essential processes that support the growth, vitality, and robustness of aquatic organisms. They serve as managers in a factory, overseeing the production of crucial proteins for muscular development and a significantly stronger immune system.

Protection and Strengthening

However, nucleotides are not just protein factory managers. In aquaculture, where disease outbreaks and environmental stresses are daily challenges, nucleotides also act as a shield for the immune system, strengthening the natural defenses of aquatic organisms and improving their ability to combat diseases more efficiently.

Nutrient Optimization

As fish and shrimp grow, their energy and nutrient demands also increase significantly, and nucleotides are indispensable for optimizing nutrient utilization, ensuring faster growth and better feed utilization, thus improving feed conversion rates.

Prosol is an Italian company proud to be one of the world’s leading producers of Ribonucleic Acid (RNA) from yeast cells using a solvent-free process. It stands out as one of the few in Europe to adopt this approach, with nucleotides produced from yeast cell RNA hydrolysis forming aqua-Care, a product specifically developed and formulated to meet the demands of key commercially important aquatic species.

Pioneering Aquatic Health and Sustainability

The art of formulating nucleotide-enriched diets has proven to be a revolutionary advancement in contemporary aquaculture. Researchers, scientists, nutritionists, and formulators are developing increasingly specific diets for different species and developmental stages. By measuring and optimizing the most appropriate levels of nucleotides in these diets, these professionals aim to enhance the growth and overall health of aquatic organisms, as well as ensure that they use their food more efficiently, reducing waste and operational costs.

“De Novo” Synthesis – The Secret to Success

However, the power of nucleotides in aquaculture goes beyond nutrition or the health and robustness of aquatic organisms. They also play a fundamental role in de novo synthesis, which we will attempt to explain in a straightforward manner.

De novo synthesis is akin to the ability to create something valuable from basic components. To illustrate, imagine that we are building something important with parts from a construction kit. In this figurative context, the parts are nucleotides, which are the essential building blocks for DNA and RNA synthesis.

In this way, nucleotide-enriched diets provide a significant amount of these fundamental building blocks to aquatic animals. Keeping the same analogy from our earlier example, it’s like offering shrimp a large supply of these “high-quality parts” to build their own DNA and RNA.

Adequate levels of nucleotides in fish and shrimp diets triggers the “de novo” synthesis of DNA and RNA, resulting in faster growth and stronger health. This approach stimulates and boosts aquaculture efficiency with clear results: healthier animals and a more profitable industry. In the next edition of ABCC Magazine, we will continue this article presenting research results that demonstrate the effectiveness of aqua-Care at different concentrations in feeds for Litopenaeus vannamei, in a challenge test with an aggressive strain of Vibrio parahemolyticus.

Article published onABCC Magazine,  FENACAM– Year XXV N. 3 – November 2023 on https://abccam.com.br/2023/11/revista-da-abcc-edicao-fenacam-ano-xxv-n-3-novembro-de-2023/

Author: Marcelo Borba, Aqua Product Manager at Prosol

Email: m.borba@prosol.it

February 2022

In addition to the existing authorisations for sows and weaned piglets, Prosol has recently received the EU authorization for its live yeast Biosprint® (MUCL 39885) now allowed as a feed additive for pig for fattening.

“We are delighted to receive this new EU authorization for Biosprint which further confirm the improvements live yeast products can have on animal performances and health – commented Emanuela Consuelo Epis, Global Product Manager at Prosol – This authorization has been achieved thanks to the dedication of the whole Prosol animal nutrition team, and we continue working so to achieve the highest quality standards and confirm results by supporting several scientific trials.”

New Prosol’s authorization was published on EU Official Journal 24/02/2022
> https://eur-lex.europa.eu/legal-content/EN/TXT/?uri=CELEX:32022R0272

Methane emissions from ruminant livestock contribute significantly to the large environmental footprint of agriculture. Ruminant production accounts for about 81% of Greenhouse Gases (GHG) from the livestock sector, 90% of which results from rumen microbial methanogenesis 1 . A considerable worldwide research effort has investigated various mitigation strategies and the ruminant farmers are putting in a lot of effort to improve
the sustainability of their operations, with reduction of methane emissions (one of the most potent greenhouse gases) being a top priority.

In ruminants, methane production is linked to fibre digestion: methanogens bacteria consume the hydrogen produced by cellulolytic bacteria. The resultant hydrogen accumulation is toxic for rumen microbiota, so methane production and eructation are important to keep the rumen healthy.

Methane production is therefore linked to dry matter intake. This means that it is crucial to carefully consider feed intake when looking to increase milk and meat production.

Several approaches have been studied in many countries with the aim of reducing methane emissions of digestive origin.  Various research has suggested that live yeasts such as Biosprint® can be used.

Biosprint is a high concentrate probiotic live yeast authorized according to European Regulation as Gut flora stabilizer Feed Additive. Biosprint®  promotes the fermentation processes, adapting the rumen environment to the needs of the bacterial flora responsible for fiber digestion. Biosprint® helps to reduce methane production by improving feed efficiency.

It has been shown that there is a negative relation between methane emission measured in g/kg of milk and daily milk production (Knapp et al., 2014).

Biosprint® can help improve daily milk production and quality without significantly increasing dry matter intake, which was demonstrated in a recent trial performed at Wageningen University.

In all the trials that we have performed we have seen an increase in milk yield, which improves the feed conversion ratio. Milk production growth is variable depending on the conditions, but it can reach up to 7%. In a trial performed at Wageningen University, Biosprint was able to improve feed efficiency (+4,2%) by increasing milk yield while decreasing body weight losses.

The extra energy extracted from the feed was partially used to increase milk production and also to reduce the negative energy balance. A better energetic status improves body condition, which can result in better fertility.

But methane is also produced by animals that do not make milk. Thus, it is important to increase the number of lactations in the herd which will help to amortise the economic and environmental cost of heifers. Biosprint® is a good support to mitigate acidosis and its sanitary consequences, like laminitis. Reduction of sanitary and metabolic problems connected to acidosis leads to a decreased culling rate, which means a lower number of heifers. Heifers do not produce milk but produce methane from their metabolism. A decreased number of heifers to maintain the herd size will help to decrease total methane production of the farm.

Biosprint® can help to reduce methane production by increasing feed efficiency in lactation and fattening, as well as by improving animal sanitary status.

Read the full Prosol interview with Feedinfo > https://bit.ly/3ipT7qq 

1. Ilma Tapio, Timothy J. Snelling, Francesco Strozzi & R. John Wallace,   The ruminal microbiome associated with methane emissions from ruminant livestock, Journal of Animal Science and Biotechnology- on   https://jasbsci.biomedcentral.com/articles/10.1186/s40104-017-0141-0
2. Feedinfo,  Live Yeast’s Impact on Rumen Microbiome Key to Industry’s Sustainability & Economic Objectives, says Prosol- INDUSTRY PERSECTIVES, 10 Jan 2022