By Dr. Ajay Bhoyar, Global Technical Manager Poultry, EW Nutrition

Gut health is a critical challenge in antibiotics-free (ABF) production as it plays a vital role in the overall health and well-being of animals. Antibiotics have long been used as a means of preventing and treating diseases in animals, but their overuse has led to the development of antibiotic-resistant bacteria. As a result, many farmers and producers are shifting towards antibiotics-free production methods. This shift presents a significant challenge as maintaining gut health without antibiotics can be difficult. It is, however, not impossible.

One of the main challenges in antibiotics-free production is the prevention of bacterial infections in the gut. The gut microbiome plays a crucial role in the immune system and overall health of animals. When the balance of microbes in the gut is disrupted (dysbiosis), it can lead to poor nutrient absorption which subsequently results in reduced live bird performance including feed efficiency and weight gain in broiler chicken. In the absence of antibiotics, farmers and producers must rely on other methods to maintain a healthy gut microbiome.

Antibiotic reduction – a major global trend

The trend in recent years has been for poultry producers to reduce their use of antibiotics to promote public health and improve the sustainability of their operations. This has been driven by concerns about the development of antibiotic-resistant bacteria and the potential impact on human health, as well as by consumer demand for meat produced without antibiotics. Many countries now have regulations in place that limit the use of antibiotics in food and animal production.

Challenges to antibiotics-free poultry (ABF) production

1. Disease control. Antibiotic-free poultry production requires farmers to rely on alternative methods for controlling and preventing diseases, such as stepped-up biosecurity practices. This can be more labor-intensive and costly.
2. Higher mortality rates. Without antibiotics, poultry farmers may experience higher mortality rates due to disease outbreaks and other health issues. This can lead to financial losses for the farmer and a reduced supply of poultry products for consumers.
3. Feeding challenges. Antibiotic growth promotors (AGPs) are often used in feed to promote growth and prevent intestinal disease in poultry. Without AGPs, poultry producers can find alternative ways to ensure expected production performance.
4. Increased cost. Antibiotic-free poultry production can be more expensive than conventional production methods, as farmers must invest in additional housing, equipment, labor, etc.

Phasing out AGPs will likely lead to changes in the microbial profile of the intestinal tract. It is hoped that strategies such as infectious disease prevention programs and using non-antibiotic alternatives minimize possible negative consequences of antibiotic removal on poultry flocks (Yegani and Korver, 2008).

Gut health is key to overall health

A healthy gastrointestinal system is important for poultry to achieve its maximum production potential. Gut health in poultry refers to the overall well-being and functioning of the gastrointestinal tract in birds. This includes the balance of beneficial bacteria, the integrity of the gut lining, and the ability to digest and absorb nutrients. Gut health is important for maintaining the overall health and well-being of the birds. A healthy gut helps to improve feed efficiency, nutrient absorption, and the overall immunity of the birds.

The gut is host to more than 640 different species of bacteria and 20+ different hormones. It digests and absorbs the vast majority of nutrients and makes up for nearly a quarter of body energy expenditure. It is also the largest immune organ in the body (Kraehenbuhl and Neutra, 1992). Consequently, ‘gut health’ is highly complex and encompasses the macro and micro-structural integrity of the gut, the balance of the microflora, and the status of the immune system (Chot, 2009).

Poultry immunity is mediated by the gut

The gut is a critical component of the immune system, as it is the first line of defense against pathogens that enter the body through the digestive system. Chickens have a specialized immune system in the gut, known as gut-associated lymphoid tissue (GALT), which helps to identify and respond to potential pathogens. The GALT includes Peyer’s Patches, which are clusters of immune cells located in the gut wall, as well as the gut-associated lymphocytes (GALs) that are found throughout the gut. These immune cells are responsible for recognizing and responding to pathogens that enter the gut.

The gut-mediated immune response in chickens involves several different mechanisms, including the activation of immune cells, the production of antibodies, and the release of inflammatory mediators. The GALT and GALs play a crucial role in this response by identifying and responding to pathogens, as well as activating other immune cells to help fight off the infection.

The gut microbiome also plays a critical role in gut-mediated immunity in chickens. The gut microbiome is made up of a highly varied community of microorganisms, and these microorganisms can have a significant impact on the immune response. For example, certain beneficial bacteria can help to stimulate the immune response and protect the gut from pathogens.

Overall, the gut microbiome, GALT, and GALs all work together to create an environment that is hostile to pathogens while supporting the growth and health of beneficial microorganisms.

Dysbiosis/Dysbacteriosis impacts performance

Dysbiosis is an imbalance in the gut microbiota because of an intestinal disruption. Dysbacteriosis can lead to wet litter and caking issues. Prolonged contact with the caked litter can lead to pododermatitis (feet ulceration) and hock-burn, resulting in welfare issues as well as degradation of the carcass (Bailey, 2010). Apart from these issues, the major economic impact comes from reduced growth rates, FCR, and increased veterinary treatment costs. Coccidiosis infection and other enteric diseases can be aggravated when dysbiosis is prevalent. Generally, animals with dysbiosis have high concentrations of Clostridium that generate more toxins, leading to necrotic enteritis.

Fig.1: Dysbiosis – a result of challenging animal’s microbiome. Source: Charisse Petersen and June L. Round. 2014

It is believed that both non-infectious and infectious factors can play a role in dysbacteriosis (DeGussem, 2007). Any changes in feed and feed raw materials, as well as the physical quality of feed, influence the balance of the gut microbiota. There are some risk periods during poultry production when the bird will be challenged, for example during feed change, vaccination, handling, transportation, etc. During these periods, the gut microbiota can fluctuate and, in some cases, if management is sub-optimal, dysbacteriosis can occur.

Infectious agents that potentially play a role in dysbacteriosis include mycotoxins, Eimeria spp., Clostridium perfringens, and other bacteria producing toxic metabolites.

Factors affecting gut health

The factors affecting broiler gut health can be summarized as follows:

1. Feed and water quality: The form, type, and quality of feed provided to broilers can significantly impact their gut health. Consistent availability of cool and hygienic drinking water is crucial for optimum production performance.
2. Stress: Stressful conditions, such as high environmental temperatures or poor ventilation, can lead to an imbalance in the gut microbiome and an increased risk of disease.
3. Microbial exposure: Exposure to pathogens or other harmful bacteria can disrupt the gut microbiome and lead to gut health issues.
4. Immune system: A robust immune system is important for maintaining gut health, as it helps to prevent the overgrowth of harmful bacteria and promote the growth of beneficial bacteria.
5. Sanitation: Keeping the broiler environment clean and free of pathogens is crucial for maintaining gut health, as bacteria and other pathogens can easily spread and disrupt the gut microbiome.
6. Management practices: Proper management practices, such as proper feeding and watering, and litter management can help to maintain gut health and prevent gut-related issues.

Fig. 2. Key factors affecting broilers’ gut health

Key approaches for managing gut health without antibiotics

Two key approaches for managing gut health in poultry without the use of antibiotics are outstandingly successful.

Proper nutrition and management practices

Ensuring the birds have access to clean water, high-quality feed, and a stress-free environment is crucial for ABF poultry production. A balanced diet in terms of protein, energy, and essential vitamins and minerals is essential for maintaining gut health.

The environment in which birds have kept plays a major role in maintaining gut health. Proper sanitation and ventilation, as well as the right temperature and humidity, are crucial to prevent the spread of disease and infection. There is no alternative to the strict implementation of stringent biosecurity measures to prevent the spread of disease.

Early detection and treatment of diseases can help to prevent them from becoming more serious problems affecting the profitability of ABF production. It is important to keep a close eye on birds for signs of disease, such as diarrhea, reduced water, and feed consumption.

Gut health-promoting feed additives

Another approach to maintaining gut health in antibiotics-free poultry production is using gut health-supporting feed additives. A variety of gut health-supporting feed additives including phytochemicals/essential oils, organic acids, probiotics, prebiotics, exogenous enzymes, etc. in combination or alone are used in animal production. Particularly, phytogenic feed additives (PFAs) have gained interest as cost-effective feed additives with already well-established effects on improving broiler chickens’ intestinal health.

Plant secondary metabolites and essential oils (generically called phytogenics, phytochemicals, or phytomolecules) are biologically active compounds that have recently garnered interest as feed additives in poultry production, due to their capacity to improve feed efficiency by enhancing the production of digestive secretions and nutrient absorption. This helps reduce the pathogenic load in the gut, exert antioxidant properties and decrease the microbial burden on the animal’s immune status (Abdelli et al. 2021).

Plant extracts – Essential oils (EOs) /Phytomolecules

Phytochemicals are naturally occurring compounds found in plants. Many phytomolecules have been found to have antimicrobial properties, meaning they can inhibit the growth or kill microorganisms such as bacteria, viruses, and fungi. Examples of phytomolecules with antimicrobial properties include compounds found in garlic, thyme, and tea tree oil. Essential oils (EOs) are raw plant extracts (flowers, leaves, roots, fruit, etc.) whereas phytomolecules are active ingredients of essential oils or other plant materials. A phytomolecule is clearly defined as one active compound. Essential oils (EOs) are important aromatic components of herbs and spices and are used as natural alternatives for replacing antibiotic growth promoters (AGPs) in poultry feed. The beneficial effects of EOs include appetite stimulation, improvement of enzyme secretion related to food digestion, and immune response activation (Krishan and Narang, 2014).

A wide variety of herbs and spices (thyme, oregano, cinnamon, rosemary, marjoram, yarrow, garlic, ginger, green tea, black cumin, and coriander, among others), as well as EOs (from thyme, oregano, cinnamon, garlic, anise, rosemary, citruses, clove, ginger), have been used in poultry, individually or mixed, for their potential application as AGP alternatives (Gadde et al., 2017).

Fig. 3: Phytomolecule-based feed additive outperforms AGPs with improved broiler performance (42 Days field study)

One of the primary modes of action of EOs is related to their antimicrobial effects which allow for controlling potential pathogens (Mohammadi and Kim, 2018).

Fig. 4: Effectivity of phytomolecule-based feed additive (Ventar D) against enteropathogenic bacteria (MIC value in PPM)

Phytomolecules have been shown to have anti-inflammatory properties. These compounds include flavonoids, polyphenols, carotenoids, and terpenes, among others. One of the ways in which phytomolecules exhibit anti-inflammatory effects is through their ability to inhibit the activity of pro-inflammatory enzymes and molecules. For example, polyphenols have been shown to inhibit the activity of nuclear factor-kappa B (NF-kB), a transcription factor that plays a key role in regulating inflammation.

Phytomolecules also have antioxidant properties, which can help to protect cells from damage caused by reactive oxygen species (ROS) and other reactive molecules that can contribute to inflammation. Plant extracts are also proposed to be used as antioxidants in animal feed, protecting animals from oxidative damage caused by free radicals. The presence of phenolic OH groups in thymol, carvacrol, and other plant extracts act as hydrogen donors to the peroxy radicals produced during the first step in lipid oxidation, thus retarding the hydroxyl peroxide formation (Farag et al., 1989, Djeridane et al., 2006). Thymol and carvacrol are reported to inhibit lipid peroxidation (Hashemipour et.al. 2013) and have strong antioxidant activity (Yanishlieva et al., 1999).

Overall, the anti-inflammatory effects of phytomolecules are thought to be due to a combination of their ability to inhibit the activity of pro-inflammatory enzymes and molecules, their antioxidant properties, and their ability to modulate the immune system. Plant extracts (i.e. carvacrol, cinnamaldehyde, eugenol. etc.) inhibit the production of pro-inflammatory cytokines and chemokines from endotoxin-stimulated immune cells and epithelial cells (Lang et al., 2004, Lee et al., 2005, Liu et al., 2020). It has been indicated that anti-inflammatory activities may be partially mediated by blocking the NF-κB activation pathway (Lee et al., 2005).

Fig. 5: Anti-inflammatory effect of phytomolecule-based feed additive (Ventar D) – the reduced activity of inflammatory cytokines

Proper protection of EOs/Phytomolecules is key to optimum results

Several phytogenic compounds have also been shown to be largely absorbed in the upper GIT, meaning that without proper protection, the majority would not reach the lower gut where they would exert their major functions (Abdelli et al. 2021). The benefits of supplementing the broiler diet with a mixture of encapsulated EOs were higher than the tested PFA in powdered, non-protected form (Hafeez et al. 2016). Novel delivery technologies have been developed to protect PFAs from the degradation and oxidation process during feed processing and storage, ease the handling, allow a slower release, and target the lower GIT (Starčević et al. 2014). The specific protection techniques used during the commercial production of an EO/phytomolecule blend are crucial in delivering on all the objectives with remarkable consistency.

Fig. 6: Pelleting stability of phytomolecule – based feed additive (Ventar D) at high temperature and longer conditioning time

Phytomolecule blend optimizes production performance

Removal of antibiotics in poultry production can be challenging for controlling mortality and maintaining the production performance of the birds. Phytogenic feed additives have been shown to improve production performance of chicken due to their antimicrobial, anti-inflammatory, antioxidant, and digestive properties. Possible mechanisms behind improved nutrient digestibility by phytogenic feed additives (PFAs) supplementation could be attributed to the ability of these feed additives to stimulate appetite, saliva secretion, intestinal mucus production, bile acid secretion, and activity of digestive enzymes such as trypsin and amylase as well as to positively affect the intestinal morphology (Oso et al. 2019). EOs are perceived as growth promoters in poultry diets, with strong antimicrobial and anticoccidial activities (Zahi et al., 2018). PFAs have positive effects on body weight gain and FCR in chickens (Khattak et al. 2014, Zhang et el. 2009).

Fig. 7: Phytomolecule-based feed additive improved broiler FCR and mortality in field trial

Conclusion

In conclusion, managing gut health is a significant challenge in ABF broiler production that must be addressed to achieve optimal performance and welfare of the birds. The use of antibiotics as a preventative measure in broiler production has been widely used, but with the increasing demand for antibiotic-free products, alternative methods to maintain gut health must be implemented. These include using gut health-supporting feed additives, and proper management practices such as implementing biosecurity measures, maintaining optimal environmental conditions, providing adequate space and ventilation, and reducing stress. However, it is essential to note that there is no one-size-fits-all solution for gut health management in ABF broiler production. It is important to continuously monitor and assess their flock’s gut health and make adjustments as necessary. Additionally, research and development in this field should be encouraged to identify new and innovative ways to maintain gut health in ABF broiler production.

Overall, managing gut health is a critical challenge that requires a multi-faceted approach and ongoing monitoring and management. By implementing the appropriate strategies and utilizing new technologies, poultry operators can ensure the health and well-being of their flocks while meeting the growing demand for antibiotic-free products sustainably.

References:

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Bailey R. A. 2010. Intestinal microbiota and the pathogenesis of dysbacteriosis in broiler chickens. PhD thesis submitted to the University of East Anglia. Institute of Food Research, United Kingdom

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De Gussem M, “Coccidiosis in Poultry: Review on Diagnosis, Control, Prevention and Interaction with Overall Gut Health,” Proceedings of the 16th European Symposium on Poultry Nutrition, Strasbourg, 26-30 August, 2007, pp. 253-261.H.J. Dorman, S.G. Deans. Antimicrobial agents from plants: antibacterial activity of plant volatile oils. J Appl Microbiol, 88 (2000), pp. 308-316

Djeridane A., M. Yousfi M, Nadjemi B, Boutassouna D., Stocker P., Vidal N. Antioxidant activity of some Algerian medicinal plants extracts containing phenolic compounds. Food Chem, 97 (2006), pp. 654-660

Farag R. S., Daw Z.Y., Hewedi F.M., El-Baroty G.S.A. Antimicrobial activity of some Egyptian spice essential oils. J Food Prot, 52 (1989), pp. 665-667

Gadde U., Kim W.H., Oh S.T., Lillehoj H.S. Alternatives to antibiotics for maximizing growth performance and feed efficiency in poultry: A review. Anim. Health Res. Rev. 2017;18:26–45.

Guo, F.C., Kwakkel, R.P., Williams, B.A., Li, W.K., Li, H.S., Luo, J.Y., Li, X.P., Wei, Y.X., Yan, Z.T. and Verstegen, M.W.A., 2004. Effects of mushroom and herb polysaccharides, as alternatives for an antibiotic, on growth performance of broilers. British Poultry Science, 45(5), pp.684-694.

Hafeez A., Männer K., Schieder C., Zentek J. Effect of supplementation of phytogenic feed additives (powdered vs. encapsulated) on performance and nutrient digestibility in broiler chickens. Poult. Sci. 2016;95:622–629.

Hammer K.A., Carson C.F., Riley T.V. Antimicrobial activity of essential oils and other plant extracts. J Appl Microbiol, 86 (1999), pp. 985-990

Hashemipour H, Kermanshahi H, Golian A, Veldkamp T. Effect of thymol and carvacrol feed supplementation on performance, antioxidant enzyme activities, fatty acid composition, digestive enzyme activities, and immune response in broiler chickens. Poultry Science. Volume 92. Issue 8. 2013, Pp 2059-2069,

Khattak F., Ronchi A., Castelli P., Sparks N. Effects of natural blend of essential oil on growth performance, blood biochemistry, cecal morphology, and carcass quality of broiler chickens. Poult. Sci. 2014;93:132–137

Kraehenbuhl, J.P. & Neutra, M.R. (1992) Molecular and cellular basis of immune protection of mucosal surfaces. Physiology Reviews, 72: 853–879.Krishan and Narang J. Adv. Vet. Anim. Res., 1(4): 156-162, December 2014

Lang A., Lahav M., Sakhnini E, Barshack I., Fidder H. H., Avidan B. Allicin inhibits spontaneous and TNF-alpha induced secretion of proinflammatory cytokines and chemokines from intestinal epithelial cells. Clin Nutr, 23 (2004), pp. 1199-1208

Lee S.H., Lee S.Y., Son D.J., Lee H., Yoo H.S., Song S. Inhibitory effect of 2′-hydroxycinnamaldehyde on nitric oxide production through inhibition of NF-kappa B activation in RAW 264.7 cells Biochem Pharmacol, 69 (2005), pp. 791-799

Liu, S., Song, M., Yun, W., Lee, J., Kim, H. and Cho, J., 2020. Effect of carvacrol essential oils on growth performance and intestinal barrier function in broilers with lipopolysaccharide challenge. Animal Production Science, 60(4), pp.545-552.

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By Lea Poppe, Regional Technical Manager, EW Nutrition

Diarrhea due to infestation with cryptosporidia is one of the most pressing problems in calf rearing. These protozoa, along with rotaviruses, are now considered the most common pathogens in infectious calf diarrhea. Due to their high resistance and thus limited possible control and prevention measures, they have now overtaken other pathogens such as coronaviruses, salmonellae, and E. coli.

Cryptosporidia show complex development

Cryptosporidia are single-celled intestinal parasites. In calves, Cryptosporidium parvum and Cryptosporidium bovis are most commonly found. C. bovis is normally considered nonpathogenic. Accordingly, the disease known as cryptosporidiosis is caused by C. parvum. The rapid tests for determining the diarrheal pathogens, which are increasingly widespread, are usually unsuitable for distinguishing between the individual strains, which can lead to false positive results.

Resistant in the environment, active in the animal

In the environment, cryptosporidia are distributed as oocysts. The oocysts are only about 5 µm in size and have a very resistant shell. They can remain infectious for up to 6 months in high humidity and moderate temperatures. Drought and extreme temperatures (below -18°C and above 65°C) cause the oocysts to die.

After oral ingestion, the oocysts are reactivated by conditions in the gastrointestinal tract (low pH and body temperature): As sporozoites, the parasites attach to the posterior small intestine, causing diarrhea symptomatology. There, they surround themselves with a special protective membrane, and the complex life cycle continues. Only a few days after infection, reproductive forms are detectable in the calf’s intestine, and excretion of infectious oocysts in the feces begins.

Figure 1 (Olias et al., 2018): Life cycle of cryptosporidia: ingested oocysts release four sporozoites that invade host enterocytes (intestinal epithelial cells). There, they develop into trophozoites before asexual and sexual reproduction ensues, and thin- and thick-walled oocysts are formed. Thick-walled oocysts are excreted through the intestine. Thin-walled oocysts may break apart, and the sporozoites may infect other enterocytes, resulting in relapse or prolonged diarrhea. Infestation of the cells leads to their destruction, resulting in villi atrophy or fusion.

Oocysts bring the disease to the animal

Cryptosporidiosis is transmitted either by direct contact of calves with feces from infected animals or indirectly by ingesting contaminated feed, bedding, or water. Each gram of feces excreted by calves showing symptoms may contain up to 100 million oocysts. According to experimental studies, as few as 17 orally ingested oocysts are sufficient to trigger infection. In addition, some multiplication forms can infect other intestinal cells directly within the intestine and thus further advance the disease by autoinfection.

Cryptosporidiosis caused by cryptosporidia often presents with typical diarrhea symptoms and occurs primarily in calves up to 3 weeks of age. Older calves may also be infected with cryptosporidia but usually show no symptoms. Pathogen excretion and, thus, the spread of disease within the herd is nevertheless likely due to the minimal infectious dose.

Damage to the intestinal wall leads to retardation of growth

Attachment of cryptosporidia to the intestinal wall is associated with an inflammatory reaction, regression and fusion of the intestinal villi, and damage to the microvilli. As a result, nutrient absorption in the small intestine is impaired, and more undigested nutrients enter the colon. The microflora starts a fermentation process with lactose and starch, leading to increased lactate levels in the blood and, thus, hyperacidity in the calf. Faintness, unwillingness to drink, recumbency, and growth disorders are the consequences.

Diarrhea often occurs late or not at all and, accordingly, is not considered the main symptom of cryptosporidiosis. When diarrhea occurs, it lasts about 1-2 weeks. The feces are typically watery, greenish-yellow, and are often described as foul-smelling. Due to diarrhea, there is a loss of electrolytes and dehydration.

Studies show: Cryptosporidia are the most prevalent diarrheal pathogens

Several studies in different regions, which examined calf diarrhea and its triggers in more detail, came to a similar conclusion: Cryptosporidia are one of the most common causes of calf diarrhea. In addition, mixed infections often occur.

DL = days of life WL = weeks of life n.m. = not mentioned  EP = enteropathogen

Cryptosporidia reduces profit

Infection with cryptosporidia and sometimes subsequent diarrhea entails treatment of the animals and generates costs (veterinarian, medication, electrolyte drinks). In addition, poorer feed conversion, lower growth, and animal losses result in lower production efficiency.

A Scottish study shows 34 kg less gain in the first six months of life compared to healthy calves in beef calves that experienced severe cryptosporidiosis in the first three weeks of life. Similar results are described in lambs, also a susceptible species to cryptosporidia. These studies suggest a long-term negative effect of cryptosporidia on growth performance and production efficiency.

Here’s how you can support your calves against cryptosporidia

High resistance of the pathogens to environmental influences, a very low necessary infection dose combined with an elevated excretion of infectious oocysts, and the possibility of autoinfection make cryptosporidia tough opponents. This is also reflected in their worldwide distribution.

What is the treatment?

Suitable drugs for the treatment of cryptosporidiosis are currently unavailable on the market. The only medicine that can be used in case of cryptosporidiosis infestation may only be administered to calves that have had diarrhea symptoms for 24 hours or less. Accordingly, this agent is usually used only for prevention. Scientific studies on its effectiveness are contradictory; some suggest that it merely delays the onset of the disease. In addition, it is not always easy to use due to the exact dosage that must be followed. Doubling the dose (sometimes happening already due to incorrectly observed intervals between doses) can lead to a toxic overdose.

Accordingly, only the symptoms of the disease – diarrhea with its accompanying symptoms – can be treated. Electrolyte and water losses must be continuously compensated with the help of a high-quality electrolyte drink. The buffer substances contained also reduce the hyperacidity of the blood caused by faulty fermentation in the intestines. For successful treatment, the electrolyte drink should be given in addition to the milk drink. Under no circumstances should the feeding of milk or milk replacer be discontinued because the sick calf urgently needs energy and nutrients. Opinions to the contrary are outdated.

As always: prevention is better than treatment

To make it more difficult for cryptosporidiosis to spread from the outset, it is worth looking at the risk factors. These include direct contact with other calves and general herd size. Furthermore, organic farms seem to have more problems with cryptosporidia. Weather also influences calves born during warmer and, at the same time, wetter weather periods (temperature-humidity index) often get sick.

Due to the limited possibilities for treatment, prevention is of greater importance. For other diarrheal pathogens such as rotavirus, coronavirus, and E. coli, it has become established practice to vaccinate dams to achieve better passive immunization of the calf. However, commercial vaccination against cryptosporidia is not currently available, making dam vaccination as unavailable as calf vaccination.

Accordingly, optimal colostrum management is the first way to protect the calf from cryptosporidia infection. This also confirms the general discussion on the Failure of Passive Transfer: various studies suggest that calves with poor immunoglobulin supply suffer from diarrhea more frequently than calves with good supply, although a concrete link to cryptosporidia itself cannot always be established with certainty.

Furthermore, it is essential to break the chain of infection within farms. In addition to the separate housing of the calves, it is necessary to ensure consistent hygiene. One should take advantage of the pathogen’s weakness as well as its sensitivity to high temperatures and ensure that the water temperature is sufficiently high when cleaning the calf pens and calving area. When disinfecting afterward, it is crucial to consider the spectrum of activity of the agent used, as not all are effective against cryptosporidia.

Egg immunoglobulins support animals against cryptosporidia

Egg immunoglobulins were initially designed to help chicks get started. In this process, hens form antibodies against pathogens they are confronted with. As studies have shown, this also works with cryptosporidia. Cama and Sterling (1991) tested their produced antibodies in the neonatal mouse model and achieved a significant (P≤0.001) reduction in parasites there. Kobayashi et al. (2004) registered decreased binding of sporozoites to the intestinal cell model and their decreased viability in addition to oocyst reduction.

In the IRIG Research Institute (2009, unpublished), feeding egg powder with immunoglobulins against cryptosporidia (10 g/day) to 15 calves reduced oocyst excretion. Before administration, calves excreted an average of 106.42 oocysts/g of feces. After administration of egg powder, only two calves still showed 103.21 oocysts/g feces, and the other 13 of the 15 calves showed no oocyst excretion. All these results are confirmed by positive customer feedback on IgY-based feed supplements.

Egg immunoglobulins and optimal colostrum management as a key solution

Since there are no effective drugs against cryptosporidia, animals must be prophylactically protected against this disease as much as possible. In addition to optimal colostrum management, which means feeding high-quality colostrum (IgG≥50g/L) to the calf as soon as possible after birth, we have products with egg immunoglobulins available to support the calf as a prophylactic against cryptosporidia infestation and thus prevent significant performance losses, especially during rearing.

References

Brainard, J., Hooper, L., McFariane, S., Hammer, C. C., Hunter, P. R., & Tyler, K. (2020). Systemic review of modifiable risk factors shows little evidential support for most current practices in Cryptosporidium management in bovine calves. Parasitology research 119, 3572-3584.

Cama, V. A., and C. R. Sterling. “Hyperimmune Hens as a Novel Source of Anti-Cryptosporidium Antibodies Suitable for Passive Immune Transfer.” University of Arizona. Wiley-Blackwell, January 1, 1991. https://experts.arizona.edu/en/publications/hyperimmune-hens-as-a-novel-source-of-anti-cryptosporidium-antibo.

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Lamp, D. O. (25. Januar 2020). Rinder aktuell: Kälberdurchfall durch Kryptosporidien – Hartnäckig und weitverbreitet. BAUERNBLATT, S. 52-53.

Losand, B., Falkenberg, U., Krömker, V., Konow, M., & Flor, J. (2. März 2021). Kälberaufzucht in MV – Alles im grünen Bereich? 30. Milchrindtag Mecklemburg-Vorpommern.

Luginbühl, A., K. Reitt, A. Metzler, M. Kollbrunner, L. Corboz, and P. Deplazes. “Feldstudie Zu Prävalenz Und Diagnostik Von Durchfallerregern Beim Neonaten Kalb Im Einzugsgebiet Einer Schweizerischen Nutztierpraxis.” Schweizer Archiv für Tierheilkunde 147, no. 6 (2005): 245–52. https://doi.org/10.1024/0036-7281.147.6.245.

Olias, P., Dettwiler, I., Hemphill, A., Deplazes, P., Steiner, A., & Meylan, M. (2018). Die Bedeutung der Cryptosporidiose für die Kälbergesundheit in der Schweiz. Schweiz Arch Tierheilkd, Band 160, Heft 6, Juni 2018, 363-374.

Santín, M., Trout, J. M., Xiao, L., Zhou, L., Greiner, E., & Fayer, R. (2004). Prevalence and age-related variation of Cryptosporidium species and genotypes in dairy calves. Veterinary Parasitology 122, 103-117.

Shaw, H. J., Innes, E. A., Marrison, L. J., Katzer, F., & Wells, B. (2020). Long-term production effects of clinical cryptosporidiosis in neonatal calves. International Journal for Parasitology 50, 371-376.

Silverlås, C., H. Bosaeus-Reineck, K. Näslund, and C. Björkman. “Is There a Need for Improved Cryptosporidium Diagnostics in Swedish Calves?” International Journal for Parasitology 43, no. 2 (2013): 155–61. https://doi.org/10.1016/j.ijpara.2012.10.009.

Thomson, Sarah, Carly A. Hamilton, Jayne C. Hope, Frank Katzer, Neil A. Mabbott, Liam J. Morrison, and Elisabeth A. Innes. “Bovine Cryptosporidiosis: Impact, Host-Parasite Interaction, and Control Strategies.” Veterinary Research 48, no. 1 (2017). https://doi.org/10.1186/s13567-017-0447-0.

Uhde, F., Kaufmann, T., Sager, H., Albini, S., Zanoni, R., & Schelling, E. (2008). Prevalence of four enteropathogens in the feces of young diarrhoeic dairy calves in Switzerland. Veterinary Record (163), 362-366.

By Dr. Inge Heinzl, Editor, EW Nutrition

Nowadays, intensive livestock farming with high stocking densities causes stress in the animals and affects the immune system^{9, 13}. The increase in respiratory diseases with associated losses and costs is only one of the consequences. Due to antimicrobial resistance, antibiotics should only be used in critical cases, so effective alternatives are requested to support the animals.

Respiratory problems are a conjunction of several factors

It already has a name: PRDC or the Porcine Respiratory Disease Complex describes the cooperation of viruses, bacteria, and non-infectious factors such as environmental conditions (e.g., insufficient ventilation), stocking density, management (e.g., all-in-all-out only by pens and not for the whole house) and pig-specific factors such as age and genetics, altogether causing respiratory issues in pigs. Non-infectious factors such as high ammonia levels weaken the immune system and lay the foundation for, e.g., mycoplasmas which damage the ciliated epithelial cells in the upper respiratory tract, the first line of defense, and pave the way for PRRS viruses. They, on their part, enter the respiratory tract embedded in inhaled dust. There, they harm the macrophages and breach a further barrier of defense. Another pathfinder is the Porcine Circovirus 2 (PCV2), which destroys specific immune cells and leads to a generally higher susceptibility to infectious agents. Bacteria such as Pasteurella multocida or Streptococcus suis further on can cause secondary infections^{7, 20, 22}. Also, the combination of mycoplasma hyopneumoniae and porcine circovirus, both typically low pathogenic organisms, leads to severe respiratory disease¹⁵.

Restricted respiratory function impacts growth

The main tasks of the respiratory tract are to take in oxygen from the air and to pump out the CO₂ entailed by the catabolism of the tissue. In pigs, however, the respiratory tract is also responsible for thermoregulation, as pigs don’t have perspiration glands. The animals must get rid of excessive heat by rapid breathing. If the respiratory function is affected due to disease, thermoregulatory capacity is reduced. The resulting lower feed intake leads to decreased growth performance and less economic profit¹⁷. One of the first studies concerning this topic was conducted by Straw et al. (1989)²¹. They asserted that, with every 10 % more affected lung tissue, daily gain decreased by about 37g. This negative correlation between affected lung tissue and weight gain could be confirmed by Paz-Sánchez et al. (2021)¹⁸. They saw that animals with >10% lung parenchyma impacted by cranioventral bronchopneumonia needed a longer time to market (208.8 days vs. 200.8 days in the control), showed a lower carcass weight (74.1 kg vs. 77.7 kg in the control group) and, therefore, also a lower daily gain (500.8 g/day compared to 567.2 g/d). In another study, Pagot and co-workers (2007)¹⁶ observed 7000 pigs from 14 French farms. They saw a significant negative correlation (p<0.001) between the prevalence of pneumonia and growth and a weight gain loss of about 0.7 for each point of pneumonia increase.

Plant extracts support pigs with different modes of action

People have always used herbal substances to cure illnesses, be it willow bark for pain, chamomile for anti-inflammation or an upset stomach. Ribwort and thyme are used as cough suppressants, and eucalyptus and menthol help you breathe better. What is good for humans can also be used for pigs. To use plant extracts efficiently, it is crucial to know their specific modes of action. Due to their volatile nature, essential oils can directly reach the target site, the respiratory tract, via inhalation¹.

1.   Plant extracts can act as an antimicrobial

Many essential oils show some degree of antimicrobial activity. So, the oils of, e. g., oregano, tea tree, lemongrass, lemon myrtle, and clove are effective against a wide range of gram-positive and gram-negative bacteria. LeBel et al. (2019)¹² tested nine different oils against microorganisms causing respiratory issues in pigs. They found the oils of cinnamon, thyme, and winter savory the most effective against Streptococcus suis, Actinobacillus pleuropneumoniae, Actinobacillus suis, Bordetella bronchiseptica, Haemophilus parasuis, and Pasteurella multocida, with MICs and MBCs from 0.01 to 0.156%.

Not only the direct bactericidal effect is important. 1,8 cineol, e.g., although often considered to have only marginal or no antimicrobial activity¹⁰, effectively causes leakage of bacterial membranes² and allows other harmful substances to enter the bacterial cell. However, cineol possesses noted antiviral properties.

2.  Plant extracts can have mucolytic, spasmolytic, and antitussive effects

In the case of respiratory disease, mucolytic and spasmolytic characteristics of phytomolecules are decisive in allowing efficient respiration. Mucolytic substances dissolve the mucus, make it more liquid and facilitate the removal from the respiratory tract by the ciliated epithelium. As liquifying the mucus with essential oils or phytomolecules is related to local irritation, dosage and application form are of the highest importance⁵.

The “cleanup” is called mucociliary clearance. There are also substances that do not dissolve the mucus but stimulate the mucociliary apparatus itself and increase mucociliary transport velocity¹.

Spasmolytic activity on airway smooth muscle is shown, for example, by menthol⁸ or the essential oil of eucalyptus tereticornis⁴. Menthol showed antitussive effects¹¹.

3.   Plant extracts can have immune-modulatory and anti-inflammatory effects

If animals are suffering from a respiratory disease or are in danger of catching one, a supportive influence on the immune system is helpful. One thing is to make vaccination more effective. Mieres-Castro et al. (2021)¹⁴ figured out that the combined application of influenza vaccine and cineol to mice resulted in a longer survival time, less inflammation, less weight loss, a lower mortality rate, less pulmonary edema, and lower viral titers after a challenge with the virus seven days after the vaccination than the mice without cineol.

On the other hand, if the animals are already ill, strengthening their immune defense is essential. Li et al. (2012)¹³ showed that interleukin-6 concentration was lower (p<0.05) and the tumor necrosis factor-α level was higher (p<0.05) in the plasma of pigs fed a diet with 0.18% thymol and cinnamaldehyde than in the negative control group. Also, the lymphocyte proliferation for pigs fed the diet with thymol and cinnamaldehyde increased significantly compared with the negative control (p<0.05).

4.   Plant extracts can act as an antioxidant

There are respiratory diseases in which reactive oxygen species (ROS) play an important role. In these cases, the antioxidant activity of phytomolecules is of interest. Here again, Li et al. (2012)¹³ asserted that a diet with 0.18% thymol and cinnamaldehyde increased the total antioxidant capacity level (p<0.05) in pigs compared to a negative control group.

Can Baser & Buchbauer (2010) described eucalyptus oil containing 1,8-cineole, the monoterpene hydrocarbons α-pinene (10–12%), p-cymene, and α-terpinene, and the monoterpene alcohol linalool, is used to treat diseases of the respiratory tract in which ROS play an important role.

5.   Plant extracts reduce the production of ammonia

High concentration of ammonia in the pig house stresses the pigs’ respiratory tract and makes them susceptible to disease. Ammonia develops when feces and urine merge and the enzyme urease degrades them. Yucca extract, containing a high percentage of saponins, can reduce ammonia emissions in animal houses. Ehrlinger (2007)⁵ supposes that the glyco-components of the saponins bind ammonia and other harmful gases. Another explanation can be the decreased activity of urease shown in a trial with rats¹⁹ or the reduction of total nitrogen, urea nitrogen, and ammonia nitrogen in sow manure³.

6.   Plant extracts often show diverse modes of useful action against respiratory issues

Due to their natural task – protecting the plant – essential oils typically do not show only one beneficial activity for us. Camphene, for example, in Thymus vulgaris, shows expectorant, spasmolytic, and antimicrobial properties and is used in treating respiratory tract infections. Menthol can be effectively used in cases of asthma due to its bronchodilatory activity on smooth muscle, its interaction with cold receptors, and the respiratory drive. Menthol acts antitussive in low concentration, gives the impression of decongestion and reduces respiratory discomfort and sensations of dyspnea.

Cineol, on its part, acts antimicrobial, antitussive, bronchodilatory, mucolytic, and anti-inflammatory. It promotes ciliary transport and improves lung function^{1, 6}. Mucolytic, antioxidant, antiviral, and antibacterial activity is ascribed to thymol⁵.

Trial shows: phytomolecules help to keep respiratory diseases in check

A field study was conducted on a Philippine piglet farm with a history of chronic respiratory issues during the growing phase, with a morbidity of about 10-15%. In this study, a supplement for water containing phytomolecules that support animals against respiratory diseases (Grippozon) was tested. For the trial, 360 randomly selected 28-day-old pigs (average weight: 6.64±0.44 kg) were divided into two groups with 6 replications per group and 30 piglets per replication. All piglets came from sows raised antibiotic-free, and the piglets received antibiotics neither upon weaning except in case of symptoms (scouring: Baytril-1 mL/pig;  respiratory disease: Excede – 1mL/pig). All piglets received the same feed and a regular water therapy regimen:

Control group: no additional supplements
Grippozon group:  Addition of 250 mL of Grippozon per 1000 L of water

As parameters, the incidence of respiratory disease, final weight, daily gain, FCR, and antibiotic cost, were recorded.

The phytomolecules-containing product reduced the incidence of respiratory diseases by 52 %, leading to a 53% lower cost for antibiotic treatment. The animals showed better growth performance (600 g higher average weight and 13 g higher average daily gain), altogether resulting in an extra cost-benefit of 1.76 US$ per pig.

Reduction in disease and medication ensures healthier pigs in the Grippozon-supplemented group, reflected by better performance.

We have means at hand to reduce the use of antibiotics

Respiratory disease is a big problem in pigs. Due to the still high occurrence of antimicrobial resistance, it is essential to reduce antibiotic use as much as possible. Phytomolecules offer the possibility to strengthen the animals’ health so that they are less susceptible to disease or support them when they are already infected. With the help of phytomolecules, we can reduce antibiotic treatments and help keep antibiotics effective when their use is indispensable.

References

1. Can Baser , K. Hüsnü, and Gerhard Buchbauer. Handbook of Essential Oils: Science, Technology, and Applications. Boca Raton, FL: Taylor & Francis distributor, 2010.
2. Carson, Christine F., Brian J. Mee, and Thomas V. Riley. “Mechanism of Action of Melaleuca Alternifolia (Tea Tree) Oil on Staphylococcus Aureus Determined by Time-Kill, Lysis, Leakage, and Salt Tolerance Assays and Electron Microscopy.” Antimicrobial Agents and Chemotherapy 46, no. 6 (2002): 1914–20. https://doi.org/10.1128/aac.46.6.1914-1920.2002.
3. Chen, Fang, Yantao Lv, Pengwei Zhu, Chang Cui, Caichi Wu, Jun Chen, Shihai Zhang, and Wutai Guan. “Dietary Yucca Schidigera Extract Supplementation during Late Gestating and Lactating Sows Improves Animal Performance, Nutrient Digestibility, and Manure Ammonia Emission.” Frontiers in Veterinary Science 8 (2021). https://doi.org/10.3389/fvets.2021.676324.
4. Coelho-de-Souza, Lívia Noronha, José Henrique Leal-Cardoso, Francisco José de Abreu Matos, Saad Lahlou, and Pedro Jorge Magalhães. “Relaxant Effects of the Essential Oil of Eucalyptus Tereticornisand Its Main Constituent 1,8-Cineole on Guinea-Pig Tracheal Smooth Muscle.” Planta Medica 71, no. 12 (2005): 1173–75. https://doi.org/10.1055/s-2005-873173.
5. Ehrlinger, Miriam. “Phytogene Zusatzstoffe in der Tierernährung.” Dissertation, Tierärztliche Fakultät LMU, 2007.
6. Gelbe Liste Online. “Gelbe Liste Pharmindex Online.” Gelbe Liste. Accessed January 20, 2023. https://www.gelbe-liste.de/.
7. Hennig-Pauka, Isabell. “Atemwegserkrankungen: Schutz fängt schon bei Ferkeln an.” Der Hoftierarzt, January 13, 2021. https://derhoftierarzt.de/2021/01/atemwegserkrankungen-schutz-faengt-schon-bei-ferkeln-an/.
8. Ito, Satoru, Hiroaki Kume, Akira Shiraki, Masashi Kondo, Yasushi Makino, Kaichiro Kamiya, and Yoshinori Hasegawa. “Inhibition by the Cold Receptor Agonists Menthol and ICILIN of Airway Smooth Muscle Contraction.” Pulmonary Pharmacology & Therapeutics 21, no. 5 (2008): 812–17. https://doi.org/10.1016/j.pupt.2008.07.001.
9. Kim, K.H., E.S. Cho, K.S. Kim, J.E. Kim, K.H. Seol, S.J. Sa, Y.M. Kim, and Y.H. Kim. “Effects of Stocking Density on Growth Performance, Carcass Grade and Immunity of Pigs Housed in Sawdust Fermentative Pigsties.” South African Journal of Animal Science 46, no. 3 (2016): 294–301. https://doi.org/10.4314/sajas.v46i3.9.
10. Kotan, Recep, Saban Kordali, and Ahmet Cakir. “Screening of Antibacterial Activities of Twenty-One Oxygenated Monoterpenes.” Zeitschrift für Naturforschung C 62, no. 7-8 (2007): 507–13. https://doi.org/10.1515/znc-2007-7-808.
11. Laude, E.A., A.H. Morice, and T.J. Grattan. “The Antitussive Effects of Menthol, Camphor, and Cineole in Conscious Guinea-Pigs.” Pulmonary Pharmacology 7, no. 3 (1994): 179–84. https://doi.org/10.1006/pulp.1994.1021.
12. LeBel, Geneviève, Katy Vaillancourt, Philippe Bercier, and Daniel Grenier. “Antibacterial Activity against Porcine Respiratory Bacterial Pathogens and in Vitro Biocompatibility of Essential Oils.” Archives of Microbiology 201, no. 6 (2019): 833–40. https://doi.org/10.1007/s00203-019-01655-7.
13. Li, Xue, Xia Xiong, Xin Wu, Gang Liu, Kai Zhou, and Yulong Yin. “Effects of Stocking Density on Growth Performance, Blood Parameters and Immunity of Growing Pigs.” Animal Nutrition 6, no. 4 (2020): 529–34. https://doi.org/10.1016/j.aninu.2020.04.001.
14. Mieres-Castro, Daniel, Sunny Ahmar, Rubab Shabbir, and Freddy Mora-Poblete. “Antiviral Activities of Eucalyptus Essential Oils: Their Effectiveness as Therapeutic Targets against Human Viruses.” Pharmaceuticals 14, no. 12 (2021): 1210. https://doi.org/10.3390/ph14121210.
15. Opriessnig, T., L. G. Giménez-Lirola, and P. G. Halbur. “Polymicrobial Respiratory Disease in Pigs.” Animal Health Research Reviews 12, no. 2 (2011): 133–48. https://doi.org/10.1017/s1466252311000120.
16. Pagot, E., P. Keita, and A. Pommier. “Relationship between Growth during the Fattening Period and Lung Lesions at Slaughter in Swine.” Revue Méd. Vét., , , 5, 253-259 158, no. 5 (2007): 253–59.
17. Pallarés Martínez, Francisco José, Jaime Gómez Laguna, Inés Ruedas Torres, José María Sánchez Carvajal, Fernanda Isabel Larenas Muñoz, Irene Magdalena Rodríguez-Gómez, and Librado Carrasco Otero. “The Economic Impact of Pneumonia Processes in Pigs.” https://www.pig333.com. Pig333.com Professional Pig Community, December 14, 2020. https://www.pig333.com/articles/the-economic-impact-of-pneumonia-processes-in-pigs_16470/.
18. Paz-Sánchez, Yania, Pedro Herráez, Óscar Quesada-Canales, Carlos G. Poveda, Josué Díaz-Delgado, María del Quintana-Montesdeoca, Elena Plamenova Stefanova, and Marisa Andrada. “Assessment of Lung Disease in Finishing Pigs at Slaughter: Pulmonary Lesions and Implications on Productivity Parameters.” Animals 11, no. 12 (2021): 3604. https://doi.org/10.3390/ani11123604.
19. Preston, R. L., S. J. Bartle, T. May, and S. R. Goodall. “Influence of Sarsaponin on Growth, Feed and Nitrogen Utilization in Growing Male Rats Fed Diets with Added Urea or Protein.” Journal of Animal Science 65, no. 2 (1987): 481–87. https://doi.org/10.2527/jas1987.652481x.
20. Ruggeri, Jessica, Cristian Salogni, Stefano Giovannini, Nicoletta Vitale, Maria Beatrice Boniotti, Attilio Corradi, Paolo Pozzi, Paolo Pasquali, and Giovanni Loris Alborali. “Association between Infectious Agents and Lesions in Post-Weaned Piglets and Fattening Heavy Pigs with Porcine Respiratory Disease Complex (PRDC).” Frontiers in Veterinary Science 7 (2020). https://doi.org/10.3389/fvets.2020.00636.
21. Straw , B. E., V. K. Tuovinen, and M. Bigras-Poulin. “Estimation of the Cost of Pneumonia in Swine Herds.” J Am Vet Med Assoc. 1989 Dec 15;195(12):1702-6. 195, no. 12 (December 15, 1989): 1702–6.
22. White, Mark. “Porcine Respiratory Disease Complex (PRDC).” Livestock 16, no. 2 (2011): 40–42. https://doi.org/10.1111/j.2044-3870.2010.00025.x.

By Dr. Ajay Bhoyar, Global Technical Manager – Poultry, EW Nutrition

Rancidity testing is essential in the feed industry, as a key indicator of product quality and shelf life. It is conducted to determine the level of oxidation in samples of feed or feed ingredients and it can be performed through a number of analytical methods.

Rancidity is the process by which fats and oils in food become degraded, resulting into off-odor/flavor, taste, and texture. This process is caused by the oxidation of unsaturated fatty acids and can be accelerated by factors such as exposure to light, heat, and air. Rancidity can occur naturally over time, but it can also be accelerated by improper storage or processing of animal products. Fats are highly susceptible to degradation due to their chemical nature.

How does oxidative rancidity occur?

Oxidation occurs when an oxygen ion replaces a hydrogen ion within a fatty acid molecule and higher numbers of double bonds within the fatty acid increase the possibility of autoxidation. Oxidative rancidity results from the breakdown of unsaturated fatty acids in the presence of oxygen. Light and heat promote this reaction, which results in the generation of aldehydes and ketones – compounds which impart off-odors and flavors to food products. Pork and chicken fat demonstrate a higher degree of unsaturated fatty acids compared with beef fat and are therefore more prone for rancidity.

Oxidation: a three-step process

Fat/oil oxidation is a three-step process (Initiation, Propagation and Termination). Therefore, the oxidation products depend on the time. In the first phase, called Initiation, the formation of free radicals begins and accelerates.

Once the initial radicals have formed, the formation of other radicals proceeds rapidly in this second phase called Propagation. In this part of the process, a chain reaction of high energy molecules, which are variations of free radicals and oxygen, are formed and can react with other fatty acids. These reactions can proceed exponentially, if not controlled. Also in this phase, the rate of peroxide radical formation will reach equilibrium with the rate of decomposition to form a bell-shaped curve.

In the final phase, called Termination, the starting material has been consumed, and the peroxide radicals, as well as other radicals decompose into secondary oxidation by-products such as esters, short chain fatty acids, polymers, alcohols, ketones and aldehydes. It is these secondary oxidation by-products, which can negatively affect the growth and performance of animals.

Fig. 1: Oxidation: a three-phase series of reactions

Antioxidants preserve the quality of rendered products

Chemical antioxidants are used in the rendering industry to help preserve the quality of animal by-products. Synthetic antioxidants, such as BHA, BHT, and ethoxyquin, can help prevent the oxidation of these by-products, which can cause them to become rancid. These chemical antioxidants are added in small amounts to the raw materials prior to rendering or can be incorporated into the finished products to help extend their shelf life and maintain their nutritional value. It is important to note that the use of antioxidants in the rendering industry must be done in compliance with regulations and guidelines set forth by the FDA and other governing bodies.

Natural antioxidants like tocopherols, rosemary extract, ascorbyl palmitate, etc. are also used to prevent oxidation and maintain the freshness of rendered products, if the chemical antioxidants cannot be used.

Rancidity testing

Rancidity testing is the process of determining the level of rancidity in a product. Testing for level of rancidity is used widely as an indication of product quality and stability.

There are several methods used for rancidity testing, including:

Organoleptic rancidity testing

Oxidation of fats and oils leads to a change in taste, smell, and appearance. Organoleptic testing involves using the senses (sight, smell, taste) to determine the level of rancidity. Trained testers will examine the product for visual signs of spoilage, such as discoloration or the presence of crystals, and will also smell and taste the product to detect any off-flavors or odors.

Chemical & instrumental rancidity testing

Chemical testing involves using chemical methods to measure the level of rancidity. One common method is the peroxide value test, which measures the amount of peroxides (indicators of rancidity) in the product. Another method is the p-anisidine test, which measures the level of aldehydes (another indicator of rancidity) in the product.

Peroxide value

Peroxide Value (PV) testing determines the amount of peroxides in the lipid portion of a sample through an iodine titration reaction targeting peroxide formations. Peroxides are the initial indicators of lipid oxidation and react further to produce secondary products such as aldehydes. Because peroxide formation increases rapidly during the early stages of rancidification but subsequently diminishes over time, it is best to pair PV testing with p-Anisidine Value to obtain a fuller picture of product quality.

Fig.2: Oxidation products changes with time

p-Anisidine (p-AV)

p-AV is a determination of the amount of reactive aldehydes and ketones in the lipid portion of a sample. Both compounds can produce strong objectionable flavors and odors at relatively low levels. The compound used for this analysis (p-Anisidine) reacts readily with aldehydes and ketones and the reaction product can be measured using a colorimeter. Samples that are particularly dark may not be the most applicable for this analysis as the colorimeter may not be able to adequately measure the wavelength required.

TBARS

Thiobarbituric acid reactive substances (TBARS) are a byproduct of lipid peroxidation (i.e. as degradation products of fats). This can be detected by the TBARS assay using thiobarbituric acid as a reagent. TBA Rancidity (TBAR) also measures aldehydes (primarily malondialdehyde) created during the oxidation of lipids. This analysis is primarily useful for low-fat samples, as the whole sample can be analyzed rather than just the extracted lipids.

The Instrumental testing involves using instruments to measure the level of rancidity.

Gas chromatography

One common method is the use of a gas chromatograph, which can detect the presence of volatile compounds that indicate rancidity.

Fourier-transform infrared spectrophotometer (FTIR)

FTIR method can detect changes in the chemical makeup of the product that indicate rancidity.

Free Fatty Acids (FFA)

FFA testing determines the fatty acids that have been liberated from their triglyceride structure. A titration is performed on the extracted fat from a specific sample. The FFA content is then determined through a calculation of the amount of titrant used to reach the final result. Knowing what type of fat or fat containing product is being tested is important for this analysis to ensure that the appropriate calculation is applied. As the test does not differentiate between fatty acid types, samples with high palmitic or lauric fatty acid composition should have a different calculation factor applied so as to accurately represent the free fatty acid result.

Oxidative Stability Index (OSI)

OSI indicates how resistant a sample is to oxidation. Samples are subjected to heat while air is injected – a process which accelerates oxidation reactions. The samples are monitored, and the time required for the sample to reach an inflection point is determined. This test is useful when testing the efficacy of an antioxidant added to a product. Antioxidants should inhibit free radical propagation and thus increase a samples ability to hold up under the stressing conditions imposed by the OSI analysis. The measuring instrument, the Rancimat.

Analytical testing considerations in rendering operations

It is common to perform regular analytical testing in a rendering operation as a part of quality control and quality assurance program. There are several methods for testing rancidity in rendering operations. It is important to choose the appropriate method based on the type of product and the desired level of accuracy.

The results of rancidity testing are used to monitor and control the rendering process to prevent or minimize rancidity. This may involve adjusting processing conditions, using antioxidants, or implementing other measures to reduce oxidation.

Table. 1: Analytical testing considerations for rendering

Conclusion

Rancidity is a common problem in rendered animal products. It can have detrimental effects on both the quality and safety of the product. It is caused by the oxidation of fats and oils, leading to the formation of harmful compounds such as free radicals and hydroperoxides. The best way to prevent rancidity is through proper storage, packaging, and handling techniques, as well as the use of antioxidants to slow down the oxidation process. It is important for manufacturers and consumers to be aware of the potential for rancidity in rendered animal products and take the necessary precautions to ensure the safety and quality of the product. 

By Predrag Persak, Regional Technical Manager, EW Nutrition

In light of sustainability requirements, shortage of feed materials, and constant pressure on energy efficiency, we must rethink how we deal with all elements that impact our production. Shrinkage is one of the essential impacting elements.  

What is Shrinkage? 

In simple terms, shrinkage is the weight loss in feed or feed materials during receiving, processing, or storage. Shrinkage happens on the farm level but also in feed mills. In this article, we will focus on the latter one. Points or reasons why this happens are diverse but not unknown. Wastages, dust, pests, moisture loss, and scale deviations are some of the most important. Through time, we found efficient ways to close the doors to feather and fur pests that were stealing valuable resources and causing shrinkage. We are also good at weight control when receiving and dispatching, by thoroughly balancing the scales. But one point related to the core of feed production – and the most significant loss – is still left untackled. That is moisture loss through grinding. 

Figure 1: Points of moisture loss and addition in the feed mill 

Grinding is one central point of shrinkage

Grinding and subsequent particle size reduction is essential from many points (handling, nutritional, processing, mixing uniformity, …) and is unavoidable if we want to produce excellent feed. In the case of grinding with hammer mills, we use kinetic energy to make the hammers beat kernels to the desired size. This is a very efficient process. However, during that process, a part of kinetic energy is also transferred to thermal, increasing the temperature of the processed feed materials and resulting in the loss of one part of valuable moisture. Also, due to size reduction and enlargement of the surface, there is much more place for evaporation and moisture movement. Losses can be up to 2%. One essential parameter for high pellet quality is the particle size, but very fine grinding will result in higher shrinkage through moisture and dust losses.

Moisture is decisive  ̶  we must manage it!

The valuable moisture is needed for many reasons. One is weight. Another reason is that nutritional density for feed materials is calculated considering a certain moisture content. Additionally, moisture influences the processing parameters during the pelleting process (targeted moisture content in the conditioner should be 16-18%). Since moisture loss is unavoidable and represents the most significant part of loss or shrinkage, we must manage it. For this purpose, we must substitute lost moisture with added moisture. And in that process, we have a short time to do it properly. Usually, we don´t have enough time for so-called “soaking”. However, with the help of surfactants, the process can be speeded up.

Surf-Ace helps to keep the moisture in the feed

Surf-Ace, a liquid preservative premix for moisture optimization, which contains organic acids / organic acid salts, emulsifiers, and surfactants, helps to keep the moisture in the feed. Conditioning can be hindered by surface tension because water forms a film on the surface of the feed particles, or oil covers the particles. Surf-Ace improves water penetration and retention by decreasing surface tension. Trials show the moisture-optimizing effect of Surf-Ace.

A trial conducted in Jordan demonstrated an increase in moisture in different processing phases (feeder, heater, and the final product). It also showed better maintenance of water in the product during storage (Fig. 2).

Figure 2: Surf-Ace achieved higher moisture levels in different phases of the feed production process

Two further trials conducted in Poland and Serbia also showed that feed millers could increase moisture in the final feed by using Surf-Ace (Fig. 3).

Figure 3: Surf-Ace provided for higher moisture content in the feed

Effective surfactants minimize shrinkage in feed

Shrinkage in times of increasing costs must be minimized by all means. The feed industry offers surfactants that keep the moisture in the feed during processing and prevent at least this part of shrinkage.

Besides the financial aspect, the optimal moisture content in feed and feed materials is important to provide high feed quality, whether concerning pellet quality or percentage of nutrients. Using surfactants, therefore, not only increases profitability but also does its bit concerning sustainability.

By Dr. Ajay Awati, Global Category Manager for Gut Health and Nutrition, EW Nutrition, and Dr. Howard Simmins, InSci Associates

After 30 years of stagnating solutions, in-feed xylanase innovation has finally arrived – with a complete focus on the needs of the broiler feed industry.

It has been over 30 years since xylanase was first introduced in broiler diets in Europe. In the meantime, it has been widely used worldwide with few, if any, major improvements. While the animal feed industry evolved in terms of production landscape, feed processing technologies and use of various by-products, xylanase enzyme technology did not keep pace. In fact, it did not evolve to meet customers’ changing needs and provide that much-needed flexibility of diet formulation for a commercial nutritionist. The wait is over: new in-feed xylanase technology is about to revolutionize broiler nutrition.

Why we need innovative xylanase enzymes for broiler production

Initially, in the 1980s, xylanase was leveraged from industries unrelated to animal production into the feed business. Gut viscosity had been a continuing problem in broiler chickens fed wheat-based diets. It led to an increased risk of enteric disease, generally reducing performance. Xylanase was shown to reduce gut viscosity in wheat-based feed by breaking down soluble arabinoxylans.

As a result, the birds grew as well as if they were fed a low-viscosity corn/soya diet. An additional benefit was lower disease risks from the reduced level of anti-nutritional factors (ANFs) and the multiple negative effects of viscosity in the intestine.

In addition to reducing viscosity, xylanase augments the release in the small intestine of nutrients from previously undigested feedstuffs. The outcome has been the use of an energy matrix value for xylanase, which essentially helps diets through least-cost formulation.

These effects account for the growth of xylanase use in the monogastric feed market. Today, the penetration is above 50%.

Limitations of existing xylanase solutions

Leveraging xylanases from other industries for viscosity reduction in poultry comes with a couple of distinct limitations:

1. Most broiler diets around the globe are based on a corn-soybean formulation, which contains far higher levels of insoluble arabinoxylans than soluble arabinoxylans. In such cases, viscosity is a relatively minor issue compared to the anti-nutritional effect of insoluble arabinoxylans.
2. The reduction of gut viscosity is less relevant in other poultry sectors, such as laying hens and turkeys.

Commercial xylanases would be required to break down insoluble NSPs in which substrate activity may be limited and difficult to predict. Fiber constituents of different cereal grains used in feed are highly variable. By- and co-products derived from cereals contain even more complex fiber components, altered further by the manner of processing that the raw material has undergone.

Additionally, poultry response is highly variable:  For an individual bird, the effectiveness of xylanase depends on the enzyme’s interaction with feed in the gastrointestinal tract (GIT) of the animal, which varies depending on the species and the animal’s age. This may explain why xylanase penetration on the feed market is not as high as that of phytase.

GH10: the next-level xylanase for feed application

A xylanase for feed is required to provide multiple functionalities, of which four are essential:

1. Capacity to break down soluble and insoluble arabinoxylan across a range of typical feedstuffs
2. Rapid activity at optimal pH in the preferred section of the GIT
3. No inhibition in the presence of xylanase inhibitors
4. Comprehensive feed processing thermostability

The GH11 family of xylanases commonly used in feed does not offer these aggregated benefits. They successfully reduce soluble NSPs in wheat-based diets, hence lowering the viscosity level in the broiler GIT. However, they are less effective in the presence of insoluble NSPs in which the arabinoxylan backbone is more complex.

Why GH10 instead of GH11?

The explanation for this can be found in the 3-dimensional structure of the GH11 xylanase. The activity of GH11 xylanases requires 3 or 4 consecutive unsubstituted xylan monomers on the backbone to find an active site. That is why they are hindered by the presence of branches, or side chains, on arabinose backbones. Consequently, they are highly specific, favoring the particularly low-branching wheat backbone.

Xylanases from the GH10 family are entirely different. Although well known, they have not been used in feed yet. The GH10 xylanases require two or fewer consecutive unsubstituted xylan monomers on the backbone to find an active site. Therefore, they can act on xylose residues near branches. This results in both more and shorter xylo-oligomers than found with GH11 xylanases. In simple terms, the GH10 xylanases have a less deep cleft than the GH11 xylanases, providing greater catalytic versatility (Pollet 2010).

Significantly, this potentially allows a broader range of feedstuffs to be incorporated into the complete diet, including co- and by-products, while maintaining performance. Therefore, with GH10, higher levels of cheaper ingredients may be included, with a significant value proposition of further reducing feed costs.

GH10 xylanases generate a range of important prebiotics

As early as 1995, it was proposed that xylanase may affect microbial activity in the gastrointestinal tract through the provision of fermentable oligosaccharides and low molecular weight polysaccharides. These are produced from the hydrolysis of soluble and insoluble arabinoxylans in cereals.

A development of particular interest is that the GH10 xylanases break down the backbone of different fibre components into small xylooligosaccharides (XOS) and arabino-xylanoligosaccharides (AXOS). This action, research shows, has value in supporting the selective growth of fibre-degrading bacteria in the large intestine, conferring positive effects on the host’s health.

The most well-known probiotic strains belong to bifidobacteria and lactobacilli, which have quite different XOS and AXOS utilization systems. Bifidobacterium adolescentis has been shown to consume AXOS and XOS, whereas Lactobacillus brevis utilises only XOS. The outcome is that AXOS releases butyrate, the short-chain fatty acid, which can improve the host’s gut barrier function, as well as reduce Salmonella colonization in broilers. Alongside these health benefits, their presence may improve performance also by reducing FCR. (Courtin et al. 2008; Ribeiro et al. 2018)

As mentioned earlier, the GH10 xylanase requires only two consecutive unsubstituted xylan monomers to cleave the xylan main chain, whereas a GH11 xylanase requires 3 or 4 consecutive unsubstituted xylan monomers. Therefore, the number of potential AXOS and XOS oligomers is higher from the action of the GH10 xylanase. This results in a wider size range of oligomers. The range is valuable as the effect is spread across the large intestine, each oligomer having a different fermentation rate. Consequently, the large intestine’s microbial activity becomes saccharolytic, which potentially reduces the undesirable products of proteolytic degradation, such as phenols and cresols.

Prebiotic combinations will vary depending on the substrate available. However, there is more flexibility in breaking down insoluble NSPs across different feedstuffs using GH10 xylanase compared to GH-11 xylanase.

The future of xylanase: Reducing feed costs through flexible formulation

EW Nutrition’s GH10-based AXXESS XY xylanase, specifically developed for animal feed, has a wide-ranging activity across typical substrates, both in corn-soy and wheat-soy diets. It also allows for a greater proportion of cheaper ingredients, enabling increased flexibility in feedstuff choices and resulting in more stable feed pricing. The activity of the GH10 xylanase in producing oligomers from the breakdown of the arabinoxylan backbone also indicates that it can produce a greater number and diversity of valuable prebiotics that sustain the growth of fiber-degrading microbiota. Consequently, the metabolism of the large intestine is shifted from proteolytic to saccharolytic, which supports the animal’s general health.

The combination of these benefits from using this xylanase results in a bird with a balanced digestive system that is more robust in the face of environmental and health challenges, supporting better performance. Furthermore, this novel enzyme solution gives nutritionists a reliable tool to reduce feed costs by being flexible in diet formulation and opportunistic in using raw materials while maintaining consistency in animal performance. Especially in these times of supply problems and raw material price hikes, such advantages are invaluable.

The naturally thermostable AXXESS XY 1000G is the most advanced xylanase yet. It is a GH10 xylanase that delivers what the industry has been asking for: a fiber-degrading enzyme suited for all poultry feed.

References

Courtin, Christophe M, Katrien Swennen, Willem F Broekaert, Quirine Swennen, Johan Buyse, Eddy Decuypere, Christiaan W Michiels, Bart De Ketelaere, and Jan A Delcour. “Effects of Dietary Inclusion of Xylooligo- Saccharides, Arabinoxylooligosaccha- Rides and Soluble Arabinoxylan on the Microbial Composition of Caecal Contents of Chickens.” Journal of the Science of Food and Agriculture 88, no. 14 (2008): 2517–22. https://doi.org/10.1002/jsfa.3373.

Ribeiro, T., V. Cardoso, L.M.A. Ferreira, M.M.S. Lordelo, E. Coelho, A.S.P. Moreira, M.R.M. Domingues, M.A. Coimbra, M.R. Bedford, and C M Fontes. “Xylo-Oligosaccharides Display a Prebiotic Activity When Used to Supplement Wheat or Corn-Based Diets for Broilers.” Poultry Science 97, no. 12 (2018): 4330–41. https://doi.org/10.3382/ps/pey336.

Pollet, Annick. “Functional and Structural Analysis of Glycoside Hydrolase Family 8, 10 and 11 Xylanases with Focus on Bacillus Subtilis Xylanase A,” 2010. https://www.biw.kuleuven.be/m2s/clmt/lmcb/publications/docs/apollet

By Judith Schmidt, Product Manager On Farm Solutions

There will always be germs in barns. Yet, calves are particularly susceptible to lung viruses and bacteria that attack the respiratory systems. What can we do to prevent calf flu?

Coughing in calves is one of the most obvious signs of illness. It should be taken seriously – calves are important for the profitability of farms. Calf flu not only leads to treatment costs but also has long-term consequences, such as weak daily gains, delayed lactation, lower milk yield, reduced fertility, and increased susceptibility to other diseases.

Respiratory disease in calves: recognize the symptoms and protect their lung health

Calves are much more sensitive to respiratory diseases than many other animals. Why? One major cause is that calves are born with immature lungs. The lungs are only fully developed at about one year of age. In addition, calves generally have small lungs relative to their body size. Furthermore, the immunological gaps around the second month of life are decisive. During this phase, the number of maternal antibodies in the calf´s blood decreases, while the calf´s own immune system is still slowly building up.

Symptoms of calf flu

1) Cough

A very easy-to-recognize sign of a developing calf flu is coughing. Coughing can also be caused by changes in weather, stress, or an unsuitable barn climate, but coughing should always be monitored, and animals should be checked for other symptoms.

2) Respiratory distress

Sick calves breathe heavily and show an increased respiratory rate. Even at rest, this can be more than forty breaths per minute, ranging from a slight acceleration of breathing to severe respiratory distress and breathing through the open mouth. Mouth breathing can be the first indication of lung damage.

3) Eye and nose discharge

Calf flu not only shows its symptoms in the internal respiratory tract but also in the eyes and nose through clear, watery discharge. In later stages, bacterial infections can also cause purulent discharge. The animal’s gaze is not clear and rather “sleepy.”

4) Body posture

Calf flu often manifests itself by drooping ears or an overall low head posture, as the calves are dull and weak. They are inactive and separate themselves from the group. They also lie down and standing up is delayed.

5) Reduced water and feed intake

Due to their physical condition, animals suffering from flu tend to take in only little feed and water or do not eat and/or drink at all. The logical consequence is a weakening of the animals. In case of doubt, one should actively water and feed the animals.

Economic significance of respiratory disease in calves

Influenza in cattle and calves is a herd disease and often causes serious financial losses. Losses are caused by pronounced performance decreases, developmental disorders of the animals, and treatment costs. Significantly reduced daily gains have been demonstrated for fattening animals.

Next to diarrheal diseases, calf flu causes the highest treatment and follow-up costs for calves. A study by the Chamber of Agriculture of Lower Saxony (Germany) found that farmers had to spend between 83 and 204 euros per sick calf, depending on the severity of the disease.

4 tips to save costs and tackle calf flu with less antibiotics use

1) Offer a stable climate

Warm, damp barns, as well as overcrowded and poorly ventilated ones, weaken the calf´s defense mechanisms. Temperature fluctuations of more than 10°C between day and night also favor the development of calf flu. It is important to keep the calves’ environment free of dust and draughts. This can be achieved by adjusting the air exchange rate.

In addition, the humidity in barns without a heating system should be between 60 and 80 percent. Data loggers help to keep an eye on the climate in the barn. They make it possible to check how the outdoor climate and ventilation affect the climate conditions in the barn.

2) Hygiene-sensitive calving management

Attention should be paid to calving management. The long-term health of the animal is already predetermined in the calving pen. If several cows calve at the same time or if calving pens are not mucked out regularly, harmful germs will accumulate. In other words: if a calf is born into a dirty box, it will absorb many germs through its mucous membranes.

3) Avoid stress

It is crucial to minimize stress from causes such as transport, re-housing, feed changes, group formation, dehorning, and weaning. These events should be spaced out as far as possible and should never occur simultaneously.

4) Prevention through supplementary feed

In the winter months, when the weather is cold and damp and constantly changing, calf flu incidence skyrockets. Now, it is imperative to strengthen the calf´s respiratory tract from the beginning. EW Nutrition’s Bronchogol Liquid is a herbal concentrate that supports respiration and stabilizes the physiological defense system in the respiratory organs.

Bronchogol liquid supports young calves in stressful situations, such as critical weather transition periods (autumn-winter; winter-spring) and housing changes, and when they suffer from calf flu. The product is based on a proprietary mixture of phytomolecules. By stimulating the cilia in the respiratory tract, the phytomolecules promote the transport of mucus and facilitate expectoration.

By Judith Schmidt, Product Manager On Farm Solutions

The average pregnancy rate for dairy cows has declined over the past decades. But why is my cow not getting pregnant? Is it because of feeding? These are questions we ask ourselves when things do not quite work out with the offspring in the cowshed. Economic success in the cow barn is closely related to the successful reproduction of our cattle herd.

The maintenance and possible improvement of fertility are becoming increasingly important issues for farm productivity. Infertility is still one of the main reasons for culling on dairy farms. When farmers decide to cull a cow after a few unsuccessful inseminations, they often ask themselves whether this could not have been prevented. There is no “all-encompassing” solution for achieving an optimal fertility rate, which ultimately requires excellent management. Relevant factors include oestrus monitoring and insemination timing, genetic conditions, feeding, hygiene, and climate.

How can I tell if a cow is in heat?

A cow behaves differently than usual during oestrus. She is restless and walks around more. A cow in heat stands next to other cows – head to tail. Sie also quarrels with her herd mates or sniffs at the shame of the other cows. Fertility in cows decreases during late winter and spring; the resulting absence of clear signs of oestrus makes it difficult to recognize the right time for insemination. There are several possible causes which will be reviewed below.

Possible causes of fertility problems in dairy cows

Beta-carotene deficiency

A productive herd needs to receive an optimal mineral and trace element supply. Beta-carotene, in particular, is essential for herd fertility. But why?

Beta-carotene is an orange-yellow plant pigment whose name comes from “carrot” because of its appearance. It is also a precursor of vitamin A. Both as a precursor and as vitamin A itself, it is essential for the organism of humans and animals, particularly when it comes to the fertility of dairy cows. Besides its important function as provitamin A, beta-carotene also exerts an independent effect on the ovary. It influences the quality of the follicle and the corpus luteum. Beta-carotene also protects the corpus luteum. It promotes the synthesis of the pregnancy hormone progesterone and thus enables the fertilized egg to implant successfully in the uterine lining.

A beta-carotene deficiency can lead to the following problems:

• Smaller, not fully functional follicles
• Altered oestrus intervals
• Indistinct signs of oestrus
• Decreased corpus luteum quality

Scientific trials show how much a beta-carotene deficiency influences the fertility process. With a beta-carotene deficiency, the fertilization rate after the first insemination is only 40%, whereas with a normal beta-carotene supply, the fertilization rate is about 70%.

How do I know if my herd is deficient in beta-carotene?

The easiest way is to check the color of the fresh colostrum. If it is a deep yellow to an even orange, the cows are supplied with sufficient beta-carotene. If it looks more ivory, this is a sign of a deficiency. Of course, a poor herd fertilization rate can also indicate a deficiency. If you suspect a beta-carotene deficiency, it is best to test some blood samples from your animal or use a testing device such as a carotene photometer. With such a test kit, you can determine not only the levels in the blood but also in the colostrum and the milk.

Feeding deficiencies

Feeding plays a major role in fertility issues. Too low input rates often have a negative effect on the health of cows. Feed quality and herd management have an impact on how long the cow loses weight after calving and at what point she gains weight again. One must always keep in mind the cows’ feeding, energy balance, and nutrient supply because cows with a negative energy balance often do not show oestrus. It is also important that the silage is of high quality – poor silage inhibits fertility.

Follicle quality

The quality of the follicle is crucial for good fertility. The quality is influenced by the energy supply during the dry period and lactation during the first days. Since the follicles are already formed in the last days of gestation, a lack of energy during this period means that the maturation of the follicles – even with a better supply later on – can no longer proceed optimally and is ultimately inferior. This inevitably leads to a reduction of oestrus symptoms and minimizes the chances of successful insemination.

Prevention is key: 4 steps to improve fertility through feeding

1) Avoid stress in the feeding environment

Well-being and a high feed intake are the basis for high milk and fattening yields as well as healthy and fertile animals. Dry cows and transit cows particularly should only experience low stress. This means no overcrowding and generous feeding space, i.e., each animal should have its own feeding space. Feeding areas that are too narrow prevent the animals from eating, rank fights occur, and feed intake decreases.

Freshly lactating cows should be separated from the group. If the cows are in calving pens or calving stables, they should always have visual contact with the herd.

2) Optimize feed quality and rations

Feed quality and feeding management determine how long the cow loses weight after calving (negative energy balance) and at what point the cow gains weight again (positive energy balance). Optimal fertility performance can only happen when a positive energy balance is achieved.

The cow’s fertility performance is primarily determined by nutrient supply and feeding. At the beginning of the lactation, high-quality basic feed with a high energy concentration should be fed, as feed intake is slow to get going after calving. Nevertheless, this ration should have sufficient structure. The amounts of concentrate should be divided into several individual portions and carefully increased. For high feed intakes, fresh water should be constantly available to the animals.

3) Treat diseases early to enable feeding

Diseases that lead to a reduced appetite should be treated as early as possible. In particular, attention should be paid to healthy hooves because a cow that has pain or difficulty getting up and walking is much less likely to go to the feed table.

4) Supplement vitamins, minerals, and trace elements

The needs-based supply of vitamins, minerals, and trace elements in every performance phase is a decisive success factor for good herd fertility. A sufficient supply of trace elements, especially selenium, manganese, zinc, as well as vitamin A and beta-carotene, are important for the formation of fertility hormones and optimal insemination success. At the same time, they ensure a high colostrum quality.

EW Nutrition’s Fertilgol Bolus is a long-term bolus to support fertility. The high content of beta-carotene has a positive influence on the formation of the corpus luteum, the oestrus cycle, the quality of colostrum and sperm. The release rate of the ingredients beta-carotene, selenium, vitamin A, and other trace elements takes place over at least twenty days. Fertilgol Bolus can be used for female and male breeding animals shortly before and during the breeding or insemination period.

By Kouji Umeda, Production Director, EW Nutrition Japan

Calves are susceptible to infection by pathogens due to their immature congenital immunity. Bovine rotavirus and bovine coronavirus, pathogenic E. coli, Clostridium, Cryptosporidium, and Eimeria spp are the major pathogens of infectious diarrhea in calves less than one month of age. Bovine rotavirus, the most frequently detected in dairy and beef cattle, is responsible for approximately 40% of diarrhea cases. In addition, 60-70% of cases of diarrhea involving bovine rotavirus occur within the first two weeks of life. Symptoms include fever, anorexia, loss of energy, and acute yellow-white watery diarrhea after 12 to 36 hours post infection, which leads to dehydration and metabolic acidosis. In more severe cases, the disease can lead to death and is considered one of the most severe diarrhea-causing pathogens in newborn calves worldwide.

Rotavirus A is a major causative pathogen of diarrhea in calf

Rotaviruses belong to the family of Reoviridae and are classified into species A to J. The rotaviruses in bovines mainly belong to species A, B, and C, which are the leading infectious agents in cattle. Calf diarrhea is primarily caused by rotavirus A (RVA). This virus is transmitted orally through feces, bedding, utensils, or people contaminated with feces. Significant diarrhea caused by the virus is attributed to

• malabsorption due to the destruction of small intestinal epithelial cells and
• inhibition of water reabsorption by enterotoxin (NSP4) produced by rotaviruses.

Adult cattle and other host animals have an immune system that protects them from infection and the development of various pathogens. As RVA exists in different genotypes, the antibodies must be specifically against this genotype; otherwise, the virus-neutralizing activity, as well as protection against infection and pathogenesis, is significantly reduced.

The classic method to prevent RVA infection

Besides adequate sanitation in the production facilities, farmers try to “improve” the composition of the maternal colostrum by vaccinating the cow. For this purpose, the cows are inoculated with inactivated, previously isolated bovine RVA. However, the immunization of calves through colostrum may not be effective enough. It also may be difficult to prevent the spread of bovine RVA by barn hygiene alone due to the recent increase in the number of cattle being raised and moved from one farm to another.

Calf diarrhea feces contain G and P genotypes of bovine RVA

In general, the three most common G genotypes of bovine RVA detected in calf diarrhea are G6, G8, and G10, and the three most common P genotypes are P[1], P[5], and P[11]. Based on the results of the genotyping survey in Japan from 1987 to 2000 (Fig. 1) and the one from 2017 to 2020 (figure 2) (Animal Health Research Division of the National Institute of Agrobiological Sciences (NIAH) together with IRIG), the bovine RVA genotypes identified as prevalent and endemic in Japan in recent years were G6P[5], G6P[11], and G10P[11]. However, the percentage of genotypes detected differed among cattle breeds (Fig. 3A, Fig. 3B, Fig. 3C).

Fig.1: Genotyping results from 1987-2000

Fig.2: Genotyping results from 2017-2020

Fig. 3A:Percentage of detection in Holstein

Fig. 3B: Detection rate in crossbreeds

Fig. 3C: Detection rate in beef cattle (Wagyu)

Cow colostrum protects the calf, egg yolk the chick AND the calf

A cow provides the calf with colostrum to ensure immunoglobulin delivery (passive immunity). In poultry, hens transfer immunoglobulins to the egg yolks and pass immunoglobulins to their chicks in this way. This biological mechanism of “immune transfer to the egg yolk” in birds can be used to arbitrarily produce yolk immunoglobulin (IgY) against pathogens of enteric infections in livestock (Ikemori et al., 1992; Ikemori et al., 1997; Yokoyama et al., 1998).

For this purpose, hens must get in contact with the respective pathogens. They produce antibodies against these pathogens – which also works with non-poultry-relevant pathogens such as bovine RVA – and transfer them to the egg (⇒IgY). The eggs with accumulated high levels of useful IgY can be collected almost daily. The immunoglobulins can be fed to livestock animals such as calves to protect them in critical times.

Continuous feeding of milk formulas containing IgY allows the IgY to remain in the intestinal lumen for a long time (Nozaki et al., 2019). There, they bind to the target pathogens and prevent infection by inhibiting their attachment to and cell invasion into intestinal epithelial cells.

IgY and genotype of the virus must match

A study verified that anti-bovine RVA IgY consisting of anti-G6P[1], anti-G6P[5], and anti-G10P[11] shows broad-spectrum virus-neutralizing activity against recent field isolates. Separate trials (see table 1) demonstrated that anti-G6 genotype IgY acted best against the RVA genotypes G6P[1] and G6P[5] and showed less activity against the G10 genotype. Anti-G10P[11] IgY worked optimally against the P[11] genotypes. The trials confirmed that either the G or the P genotype must match to achieve a sufficient virus-neutralizing activity. The IgY mixture is not helpful against bovine RVA strains that match neither the G nor the P genotypes (Odagiri et., 2020).

As the genotyping survey of 2017-2020 showed mainly G6 and G10 genotypes, a mixture of anti- bovine RVA G6P[1] IgY, G6P[5], and G10P[11] has strong virus neutralizing activity against bovine RVA that is currently prevalent and spreading in production sites.

Table 1: Virus-neutralizing activity of field-isolated bovine RVA against various genotypic strains

+++：Strong virus neutralizing activity; ++：Moderate virus neutralizing activity; +：Weak virus neutralizing activity; －：No virus neutralizing activity

Anti-bovine RVA IgY supports calves against rotavirus infection

To verify the protective effect of oral passive immunization with anti-bovine RVA IgY against bovine RVA infection, a trial with newborn calves was conducted.

Trial design: Eight calves were separated from their mothers immediately after birth without feeding colostrum and moved to a house with infected animals. From the first day, the calves were fed artificial milk supplemented with anti-bovine RVA IgY (n=4) or non-immune IgY (Control IgY; n=4) three times a day.

The parameters observed were fecal score, bovine RVA excretion, and weight gain; data were collected daily. The fecal score was calculated as the cumulative fecal score during the study period: 0 for normal stools, 1 for soft to muddy stools, and 2 for watery stools. Bovine RVA was isolated from daily fecal samples and evaluated by the total number of days of bovine RVA excretion.

Results: The anti-bovine RVA IgY group was found to be effective in reducing the incidence of diarrhea and shortening the duration of virus excretion in the infection test with the bovine RVA G6 genotype strain and the bovine RVA G10 genotype strain (tables 2 and 3).

Table 2: Efficacy of anti-bovine RVA IgY feeding in bovine RVA G6 genotype strain infection

**： P＜0.01; *： P＜0.05

Table 3: Efficacy of anti-bovine RVA IgY feeding in bovine RVA G10 genotype strain infection

**： P＜0.001

IgY is a valuable tool in rotavirus control

Newborn calves, susceptible to severe diarrhea caused by bovine RVA infection, require passive immunization with antibodies transferred from the colostrum of the mother cow. However, sometimes, calves don’t get enough antibodies which can be the case if

• the calf does not receive enough colostrum or receives it too late
• the cow still has not the farm-specific antibodies because of a too short time of being on the farm

To compensate for this lack of immunity, calves have been fed milk formulas containing anti-bovine RVA IgY for some time. Continuous feeding of anti-bovine RVA IgY, which shows strong virus neutralizing activity against each genotype of bovine RVA isolated from recent cases of calf diarrhea, is expected to provide sufficient immunity and be an effective means of bovine RVA control.

In the case of disease outbreaks, it makes sense to utilize IgY with appropriate mechanisms of action in addition to improving the level of quarantine measures, including hygiene control and vaccination.

References:

Ikemori, Yutaka, Masahiko Kuroki, Robert C. Peralta, Hideaki Yokoyama, and Yoshikatsu Kodama. “Protection of Neonatal Calves against Fatal Enteric Colibacillosis by Administration of Egg Yolk Powder from Hens Immunized with K99-Piliated Enterotoxigenic Escherichia Coli.” Amer. J. Vet. Res. 53, no. 11 (1992): 2005–8. PMID: 1466492.

Ikemori, Yutaka, Masashi Ohta, Kouji Umeda, Faustino C. Icatlo, Masahiko Kuroki, Hideaki Yokoyama, and Yoshikatsu Kodama. “Passive Protection of Neonatal Calves against Bovine Coronavirus-Induced Diarrhea by Administration of Egg Yolk or Colostrum Antibody Powder.” Veterinary Microbiology 58, no. 2-4 (1997): 105–11. https://doi.org/10.1016/s0378-1135(97)00144-2.

Nozaki, I., M. Itoh, F. Murakoshi, T. Aoki, K. Shibano, and K. Yamada. “Effect of an Egg Yolk Immunoglobulin（Igy）Product on Oocyst Shedding and Blood and Fecal Igy Concentrations in Cryptosporidium-Infected Calves.” Japanese Journal of Large Animal Clinics 10, no. 2 (2019): 68–72. https://doi.org/10.4190/jjlac.10.68.

Odagiri, Koki, Nobuki Yoshizawa, Hisae Sakihara, Koji Umeda, Shofiqur Rahman, Sa Van Nguyen, and Tohru Suzuki. “Development of Genotype-Specific Anti-Bovine Rotavirus a Immunoglobulin Yolk Based on a Current Molecular Epidemiological Analysis of Bovine Rotaviruses a Collected in Japan during 2017–2020.” Viruses 12, no. 12 (2020): 1386. https://doi.org/10.3390/v12121386.

Yokoyama, Hideaki, Robert C. Peralta, Kouji Umeda, Tomomi Hashi, Faustino C. Icatlo, Masahiko Kuroki, Yutaka Ikemori, and Yoshikatsu Kodama. “Prevention of Fatal Salmonelosis in Neonatal Calves, Using Orally Administered Chicken Egg Yolk Salmonella-Specific Antibodies.” Amer. J. Vet. Res. 59, no. 4 (1998): 416–20. PMID: 9563623.

By Dr. Inge Heinzl, Editor, EW Nutrition 

For a long time now, IgY technology has been used to provide clear benefits in diagnostics, human medicine, and animal production. To give you a deeper insight into this topic, in the following, we will show you some steps of production, the benefits, and the applications of IgY.

IgY – what is it?

A short time later, Klemperer (1892) documented that the serum antibodies were also transferred into the egg. For this purpose, he did a similar trial with hens but collected the eggs. He fed mice a solution containing the egg yolk, and afterward, he infected them with tetanus. All mice with a higher dosage of egg yolk remained healthy, the others receiving a low dosage or no egg yolk died.

IgY production is a non-invasive and highly effective process

The “usual” production of antibodies in mammals includes pain and stress-causing procedures such as immunization, bleeding, and sacrifice. The only stress factor in producing egg antibodies is the hyper-immunization with the pathogen or parts of it; the rest -collecting the eggs- is non-invasive (Ikemori et al., 1993). The European Centre for the Validation of Alternative Methods (ECVAM) ), one of Europe’s health and consumer protection institutes, strongly recommends egg immunoglobulins as an alternative to mammalian antibodies (Schade et al., 1996).

IgY production is also advantageous in terms of quantitative and qualitative output. Usually, one egg (with 15 mL of yolk) contains about 100-150 mg IgY  (Pereira et al., 2019). Assuming that a hen lays about 300 eggs per year, one bird can produce between 30 and 45 g IgY in this period. After the isolation of the IgY from the egg yolk and the extraction from the remaining proteins, a final purification step that includes chromatography could achieve IgY with >90 % purity (Morgan et al., 2021).

Hyperimmunized hens provide more effective IgY

The targeted confrontation of the animal with specific pathogens or antigens leads to the production of specific antibodies. In a field trial with piglets, Kellner et al. (1994) compared three groups of piglets suffering from diarrhea on day 1 of the test. One group received egg powder originating from hens hyperimmunized with diarrhea-causing pathogens, the second group egg powder from regular eggs, and the third didn’t receive any egg powder. The following results they achieved in one of two farms. The trial shows that, after applying egg powder with selected antibodies, the animals completely recovered within three days. In the group receiving egg powder of regular eggs, still, 9.1% suffered from severe diarrhea and in the control group without any egg powder, only 27.3 % recovered.

Figure 1: Comparison of eggs originating from regular and hyperimmunized hens

Preconditions for and benefits of industrially produced IgY

A process must meet specific requirements to be suitable for industrial production. In the case of IgY production, the crucial preconditions are that…

• hens produce antibodies also against pathogens non-specific to them
• the antibodies produced and transferred to the egg also are effective in mammals (Yokoyama et al., 1993)
• due to their phylogenetic distance from mammals, hens can produce antibodies even against structurally highly conserved proteins, which is not always possible in rabbits, guinea pigs, and goats (Gassman and Hübscher, 1992).

Industrially produced IgY can target selected pathogens, e.g., enteric bacteria or viruses, respiratory pathogens, SARS-COV-2, etc. As the antibodies act not only in birds but also in other animals, such as mammals including humans, they can be used to prevent disease or support persons/animals in the case of illness. IgY is safe for animals and humans.

Concerning the economic benefits of IgY production, it can be said that it is a cost-effective method due to the high concentration of IgY in the egg yolk and the relatively simple process of the purification of the antibodies. Additionally, feeding and handling are easier and more cost-effective for hens than for many other animals.

Not all IgY products are the same

There are different methods of IgY production. One possibility is to hyperimmunize the hens simultaneously with multiple antigens. This method seems to be convenient but does not deliver standardized products concerning the content of immunoglobulins.

The other possibility is the immunization of different groups of hens, each with one antigen (e.g., Rotavirus, Salmonella, E. coli). The content of immunoglobulins is determined, and the different egg powders are mixed. The result is an IgY product with standardized amounts of specific immunoglobulins.

Where can we use IgY?

There are different application areas for IgY or IgY products. In human medicine, egg immunoglobulins can be used against the toxin of rattlesnakes or scorpions, or Streptococcus mutans bacteria, causing dental caries (Gassmann and Hübscher, 1992) Egg immunoglobulins are important for diagnostic tests such as radioimmunoassay (RIA) and enzyme-linked immunoassay (ELISA).

A further application area is animal nutrition. Young animals, such as calves or piglets, but also young dogs or cats, are born with immature immune systems. If they, additionally, are deprived of maternal colostrum in adequate quantity and/or quality, they suffer from immunity gaps during their first weeks of life and are susceptible to pathogens in their environment.

Antibiotics have been used prophylactically for a long time to protect young animals in this critical phase. With increasing antibiotic resistance, this procedure is not allowed anymore.

Products based on egg immunoglobulins against enteric pathogens, e.g., support young animals against newborn or weaning diarrhea (e.g., Yokoyama et al., 1992; Ikemori et al., 1992; Ikemori et al., 1997, Yokoyama et al., 1998).

IgY – a fascinating technology that should be better recognized

IgY technology is an animal-friendly technology with high output. Its various applications make IgY a helpful tool for human medicine as well as animal production. To get the best results, attention must be paid to quality, meaning, amongst others the standardization of the products.

IgY is an optimal tool to help young animals such as calves and piglets cope with pathogenic challenges in early life. Consequently, IgY technology enables us to limit (preventive) antimicrobial use in critical periods of animal rearing and, therefore, reduce antimicrobial resistance.

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