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Cryptosporidia in calves – chickens can help

Header Calf Standing Fotolia L

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.

Header Calf En
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.

 

Country or region Number Age/Health status % Crypto-sporidia % Rota viruses Combined infections with crypto-sporidia Others (%) Source
Switzerland 2 – 21 DL

Ill and healthy

43 46 1 case of E. coli Luginbühl et al., 2012
Switzerland 63 1 – 4 DL

Ill and healthy

————–

7 – 20 DL

————–

26 – 49 DL

34.4

—————

54.0

—————

33.3

3.1

—————-

28.6

—————-

13.3

2 KE – 1.6

4 KE – 3.2

—————-

2 KE – 19

3 KE – 3.2

4 KE – 0

—————-

2 KE – 30

3 KE – 11.7

4 KE – 6.7

Corona 4.7

E. coli 4.7

Giardia 1.6

—————

Corona 0

E. coli 3.2

Giardia 6.3

—————

Corona 0

E. coli 15

Giardia 35

 

Weber et al., 2016

 

Weber et al., 2016 EN

Switzerland 147 Up to 3rd WL;

Diarrhea

55 58.7 5.5 % Rota

7.8 % BCV

Lanz Uhde et al., 2014
Sweden 782 1 – 7 DL

Diarrhea

25.3 Detected with Giardia, E. coli, Rota, Eimeria Silverlås et al., 2012
USA (East coast) 503 Pre-weaning 50.3 Santin et al., 2004
USA 30 2 weeks old

1-8 weeks old

3-12 months

12-24 months

96.7

45.8

18.5

2.2

Santin et al., 2008
Germany 521 32 9 Losand et al., 2021
Ethiopia 360 18.6 Ayele et al., 2018
Argentina 1073 n.m. / Ill and healthy 25.5 Lombardelli et al., 2019
UK n.m. Ill ?? 37 25 20 Coccidia 8

E. coli 4

Corona 3

Co infections not including Crypto-sporidia 3

APHA, SRUC, Veterinary investigation diagnosis analysis (VIDA) report (2014)

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

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 oocytes/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.

 




Respiratory challenges in pigs: Plants to the rescue!

Swine Pig Pixabay

Nowadays, intensive livestock farming with high stocking densities causes stress in the animals and affects the immune system9, 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 infections7, 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 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 function1, 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:

Week 1 (1st week after weaning):
  • multivitamins, amino acids – 200-400 g/1000 L of water
  • water acidifier I (citric acid +enzyme) – 2 L/1000 L
Week 2-10:
  • water acidifier II (citric acid) – 300-400 mL/1000 L)

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.

Graph Phytomolecules

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.



Rancidity in fats and oils: Considerations for analytical testing

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by 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.

Three Phase
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.

Three Phase Graph
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.

Test objective Analysis Remarks
Current state of oxidation
  1. Peroxide Value (PV)
  2. Secondary Oxidatives (p-Anisidine, TBARS)
  1. PV:< 5 meq/kg
  2. 50 ppm
Potential for future oxidation Oxidative Stability Index (OSI) Analyze the stability of oil/fats
Residual antioxidant Gas chromatography Value decreases as the antioxidant gets sacrificed

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. 




IgY supports calves against rotavirus infections

Picture Header Image Dsc

By Kouji Umeda

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

IgY Virus-neutralizing test strain
SMN 1 HKD 18 SMN 35 HKD 6 HKD 7 HKD 17 KK-3 OKY 31 MYG 1 Dai-10
1978 2018 2018 2017 2017 2017 1983 2017 2017 2007
G6P[1] G6P[5] G6P[5] G6P[11] G6P[11] G6P[11] G10P[11] G10P[11] G8P[14] G24P[33]
anti-G6P[1] 1978 IgY +++ +++ +++ +++ +++ +++ + +
anti-G6P[5] 2018 IgY +++ +++ +++ ++ ++ ++ + + +
Anti-G10P[11] 2017 IgY + + + + ++ ++ +++ +++
Control IgY

+++: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

Test Group Diarrhea incidence Cumulative fecal score Bovine RVA excretion days Increase in body weight
(n animals affected/n animals tested) kg %
Anti-bovine RVA IgY     0%   (0/4) 0.0 ± 0.0* 2.3 ± 0.5** 1.3± 0.4** 3.5 ± 0.7**
Control IgY 100%  (4/4) 12.8 ± 4.8 7.8 ± 1.3 – 3.3 ± 1.6 – 7.6 ± 3.6

**: P<0.01; *: P<0.05

 

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

Test Group Diarrhea incidence Cumulative fecal score Bovine RVA excretion days increase in body weight
(n animals affected/n animals tested) kg %
Anti-bovine RVA IgY 50%   (2/4) 2.3 ± 4.5** 4.3 ± 1.3** 1.1± 0.8** 3.3 ± 3.1**
Control IgY 100%  (4/4) 14.5 ± 3.7 7.3 ± 1.0 – 4.2 ± 0.7 – 11.1 ± 2.1

**: 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.




IgY technology: using nature to support antibiotic reduction

Header Image Eggs Egg Breaking

Authors: Inge Heinzl, Lucas Queiroz

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?

IgY (immunoglobulin of the yolk) are immunoglobulins that hens produce to protect their chicks during the first weeks of life against occurring pathogens. They are the equivalent of immunoglobulin G in the colostrum of mammalians. IgY are an entirely natural product; every egg sold in the supermarket contains IgY.

IgY develops in the hen against the pathogens with which the hens are confronted. Thereby, it does not matter if these pathogens are relevant for the hens. They also produce antibodies against, e. g., bovine, porcine, or human-specific pathogens. This fact was already noticed by Vaillard (1891). He saw that the intraperitoneal injection of tetanus bacteria raised immunity against tetanus bacteria in hens’ serum.

Egg immunoglobulins

 

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 specific 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.

Comparison Of Eggs Originating From Regular And Hyperimmunized HensFigure 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 proteins structurally highly conserved, which is not always possible in rabbits, guinea pigs, and goats (Gassman and Hübscher, 1992).

Industrially produced IgY can target specific 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 and humans, they can be used to prevent disease or support persons/animals in the case of illness. If the technique is mastered, the production of IgY is not complicated. 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 several 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 defective immune equipment. Suppose they additionally don’t get maternal colostrum in adequate quantity and quality. In that case, 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 established

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

IgY technology enables us to restrict preventive antimicrobial use in critical periods of animal rearing and, therefore, reduce resistance. IgY is an optimal tool to help young animals such as calves and piglets cope with the first risky weeks of life.

References:

Gassmann, M., and U. Hübscher. “Use of Polyclonal Antibodies from Egg Yolk of Immunised Chickens .” ALTEX – Alternatives to animal experimentation 9, no. 1 (1992): 5–14.

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. https://doi.org/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.

Ikemori, Yutaka, Robert C. Peralta, Masahiko Kuroki, Hideaki Yokoyama, and Yoshikatsu Kodama. “Research Note: Avidity of Chicken Yolk Antibodies to Enterotoxigenic Escherichia Coli Fimbriae.” Poultry Science 72, no. 12 (1993): 2361–65. https://doi.org/10.3382/ps.0722361.

Kellner, J., M.H. Erhard, M. Renner, and U. Lösch. “Therapeutischer Einsatz Von Spezifischen Eiantikörpern Bei Saugferkeldurchfall – Ein Feldversuch.” Tierärztliche Umschau 49, no. 1 (January 1, 1994): 31–34.

Klemperer, Felix. “Ueber Natürliche Immunität Und Ihre Verwerthung Für Die Immunisirungstherapie.” Archiv für Experimentelle Pathologie und Pharmakologie 31, no. 4-5 (1893): 356–82. https://doi.org/10.1007/bf01832882.

Pereira, E.P.V., M.F. van Tilburg, E.O.P.T. Florean, and M.I.F. Guedes. “Egg Yolk Antibodies (Igy) and Their Applications in Human and Veterinary Health: A Review.” International Immunopharmacology 73 (2019): 293–303. https://doi.org/10.1016/j.intimp.2019.05.015.

Schade, R., C. Staak, C. Hendriksen, M. Erhard, H. Hugl, G. Koch, A. Larsson, et al. “The Production of Avian (Egg Yolk) Antibodies: IgY,” 1996. https://www.researchgate.net/publication/281466059_The_production_of_avian_egg_yolk_antibodies_IgY_The_report_and_recommendations_of_ECVAM_workshop_21.

Schade, R., C. Staak, C. Hendriksen, M. Erhard, H. Hugl, G. Koch, A. Larsson, et al. “The Production of Avian (Egg Yolk) Antibodies: IgY. The Report and Recommendations of ECVAM Workshop 21.” ATLA (Alternatives to Laboratory Animals) 24 (1996): 925–34. https://doi.org/https://www.researchgate.net/publication/281466059_The_production_of_avian_egg_yolk_antibodies_IgY_The_report_and_recommendations_of_ECVAM_workshop_21.

Yokoyama, H, R C Peralta, R Diaz, S Sendo, Y Ikemori, and Y Kodama. “Passive Protective Effect of Chicken Egg Yolk Immunoglobulins against Experimental Enterotoxigenic Escherichia Coli Infection in Neonatal Piglets.” Infection and Immunity 60, no. 3 (1992): 998–1007. https://doi.org/10.1128/iai.60.3.998-1007.1992.

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. https://doi.org/PMID: 9563623.

Yokoyama, Hideaki, Robert C. Peralta, Sadako Sendo, Yutaka Ikemori, and Yoshikatsu Kodama. “Detection of Passage and Absorption of Chicken Egg Yolk Immunoglobulins in the Gastrointestinal Tract of Pigs by Use of Enzyme-Linked Immunosorbent Assay and Fluorescent Antibody Testing.” American Journal of Veterinary Research 54, no. 6 (1993): 867–72. https://doi.org/PMID: 8323054.

Zhang, Xiao-Ying, Ricardo S. Vieira-Pires, Patricia M. Morgan, Rüdiger Schade, Xiao-Ying Zhang, Rao Wu, Shikun Ge, and Álvaro Ferreira Júnior. “Immunization of Hens.” Essay. In IGY-Technology: Production and Application of Egg Yolk Antibodies. Basic Knowledge for a Successful Practice., 116–34. Cham, Switzerland: Springer Nature, 2021.

Zhang, Xiao-Ying, Ricardo S. Vieira-Pires, Patricia M. Morgan, Schade Rüdiger, Patricia M. Morgan, Marga G. Freire, Ana Paula M. Tavares, Antonysamy Michael, and Xiao-Ying Zhang. “Extraction and Purification of IgY .” Essay. In IGY-Technology: Basic Knowledge for a Successful Practice, 135–60. Cham: Springer International Publishing AG, 2021.

Zhang, Xiao-Ying, Ricardo S. Vieira-Pires, Patricia M. Morgan, Schade Rüdiger, Patricia M. Morgan, Xiao-Ying Zhang, Antonysamy Michael, Ana Paula M. Tavares, and Marga G. Freire. “Extraction and Purification of IgY (Chapter 11).” Essay. In IGY-Technology: Basic Knowledge for a Successful Practice, 135–60. Cham: Springer International Publishing AG, 2021.




Effective phytomolecules combine superior processing stability and strong action in the animal

Phytomolecules

For millennia, plants have been used for medicinal purposes in human and veterinary medicine and as spices in the kitchen. Since the ban of antibiotic growth promoters in 2006 by the European Union, they also came into focus in animal nutrition. Due to their digestive, antimicrobial, and gut health-promoting characteristics, they seemed an ideal alternative to compensate for the reduced use of antibiotics in critical periods such as brooding, feed change or gut-related stress.

To optimize the benefits of phytomolecules, it is crucial that

  • the phytomolecules levels are standardized for consistent results and synergy
  • they show the highest stability during stringent feed processing; being often highly volatile substances, they should not get lost at high temperatures and pressure
  • the phytomolecules are preferably completely released and available in the animal to achieve the best effectiveness.

First step: Standardized phytomolecules

Essential oils and other phytogenics are sourced from plants. The composition of the plants substantially depends on genetic dissimilarity within accessions, plant origin, the site conditions, such as weather, soil, community, and harvest time, but also sample drying, storage, and extraction processes (Sadeh et al., 2019; Yang et al., 2018; Ehrlinger, 2007). For example, the oil extracted from thyme can contain between 22 and 71 % of the relevant phenol thymol (Soković et al., 2009; Shabnum and Wagay, 2011; Kowalczyk et al., 2020).

Modern technology enables the production of standardized phytomolecules with the highest degree of purity and lowest possible batch-to-batch variation for high-quality products. It also offers increased environmental and economic sustainability due to reliable and cost-effective sourcing technology.

Using such highly standardized phytomolecules enables the production of phytogenic-based feed supplements of consistently high quality.

Second step: Selection of the most suitable phytomolecules

Phytomolecules have different primary characteristics. Some support digestion (Cho et al., 2006, Oetting, 2006; Hernandez, 2004); others act against pathogens (Sienkiewitz et al., 2013; Smith-Palmer et al., 1998; Özer et al., 2007) or are antioxidants (Wei and Shibamoto, 2007; Cuppett and Hall, 1998). To optimize gut health in animal production, one of the main promising mechanisms is reducing pathogens while promoting beneficial microbes. The decrease of pathogens in the gut not only decreases the risk of enteritis incidence but also eliminates the inconvenient competitors for feed.

In order to find out the best combination serving the intended purpose, a high number of different phytomolecules need to be evaluated concerning their structure, chemical properties, and biological activities first. Availability and costs of the substances are further factors to consider. With the selection of the most suitable phytomolecules, different mixtures are produced and tested for their effectiveness. Here, it is essential to concern synergistic or antagonistic effects.

For an effective and efficient blend of phytomolecules, many steps of selection and tests are necessary – and as a result, possibly only a few mixtures can meet the requirements.

Third step: Protecting the ingredients

Many phytomolecules are inherently highly volatile. So, only having a standardized content of phytogenics in the product can not ensure the full availability of phytomolecules when used through animal feed. Some parts of the ingredients might already get lost in the feed mill due to the stringent feed hygienization process followed by feed millers to reduce pathogenic load. The heating is a significant challenge for the highly-volatile components in a phytomolecule-based product. So, protecting these phytomolecules becomes imperative to guarantee that the phytomolecules put into the feed will reach the animal.

A delicate balancing act is required to ensure the availability and activity of phytomolecules at the right site in the gut. The phytomolecules must not get lost during feed processing but must also be released in the intestine. A carrier with capillary binding of the phytomolecules together with a protective coating can be one of the available effective solutions. It protects the ingredients during feed processing, but the digestive tract’s pH and enzymes open the coating, and the phytomolecules would be available for an activity at the right site.

Study shows excellent stability of Ventar D under challenging conditions

Ventar D is a latest generation phytomolecule-based solution for gut health optimization introduced by ​EW Nutrition, GmbH. A scientific study was conducted to compare the stability of Ventar D, in the pelleting process, with two leading phytogenics competitor feed supplements.

For this trial, feed with the different added phytogenic feed supplements had to undergo a conditioning and pelletization process. The active ingredients were analyzed before and after the pelletization process. All phytogenic feed supplements under testing were added to standard broiler feed at the producer’s recommended inclusion rate. The tests took place under conditioning times of 45, 90, and 180 seconds and pelleting temperatures of 70, 80, and 90°C (158, 176, and 194°F). After cooling, triplicate samples were collected and analyzed. The respective marker substance was analyzed through gas chromatography/mass spectrometry (GC/MS) analysis to measure the recovery rate in the finished feed. The phytomolecule content of the mash feed (before pelletization) found by the laboratory was used as a baseline and set to 100% recovery. The recovery rates of the pelleted feed were evaluated relative to this baseline.

The results are presented in figure 1. Ventar D showed the highest stability of active ingredients with recovery rates of 90% at 70°C/45 sec. or 80°C/90 sec and 84% at 90°C/180 sec. The modern production technology used for Ventar D ensures that the active ingredients are well protected throughout the pelletization process.

Figure Recovery Rates Heat StabilityFigure 1: Phytomolecule stability under processing conditions, relative to mash baseline (100%)

Another trial was conducted in a feed mill in the US. For this trial, ten samples were collected from different batches of mash feed where Ventar D was added at 110g/t. Conditioning of the mash feed was at 87.8°C (190°F) for 6 minutes and 45 seconds. After the pelleting process, ten samples from the pelleted feed were collected from the continuous flow with a 5 min gap between the samplings to determine Ventar D’s recovery.

The average recovery achieved for Ventar D was 92%.

Trials show improved growth performance

Initial trials showed Ventar D’s complete release in digestion models. To examine the benefit in in-vivo conditions, Ventar D was tested in broilers at an inclusion rate of 100 g/MT.

Several in vitro studies proved the antimicrobial activity of Ventar D. One test also confirms that Ventar D could exhibit differential antimicrobial activity by having stronger activity against common enteropathogenic bacteria while sparing the beneficial ones (Heinzl, 2022). Moreover, Ventar D’s antioxidant and anti-inflammatory activity support better gut barrier functioning. Better gut health leads to higher growth performance and improved feed conversion, which could be demonstrated in several trials with broilers (figures 2 and 3). In the tests, a group fed Ventar D was compared to either a control group with no such feed supplement or groups supplied with competitor products at the recommended inclusion rates.

Compared to a negative control group, the Ventar D group consistently showed a higher average daily gain of 0.3-4.1 g (0.5-8.5 %)  and a 3-4 points better feed conversion. Compared to competitor products, Ventar D provided 1-1.7 g (2-3 %) higher average daily gain and a 3 points better /1 point higher FCR than competitors 2 and 1.

Figure Daily GainFigure 2: Average daily gain (g) – results of several trials conducted with broilers

 

Figure FcrFigure 3: FCR – results of several trials conducted with broilers

Standardization and new technologies for higher profitability

Several in vitro and in vivo studies proved that Ventar D takes “phytomolecules’ power” to the next level: Combining standardized phytomolecules and optimal active ingredient protection leads to superior product stability during feed processing. The higher amount of active ingredients arriving in the gut improves gut health and increases the production performance of the animals. Ventar D shows how we can use phytomolecules more effectively and benefit from higher farm profitability.

 

References:

Cho, J. H., Y. J. Chen, B. J. Min, H. J. Kim, O. S. Kwon, K. S. Shon, I. H. Kim, S. J. Kim, and A. Asamer. “Effects of Essential Oils Supplementation on Growth Performance, IGG Concentration and Fecal Noxious Gas Concentration of Weaned Pigs”. Asian-Australasian Journal of Animal Sciences 19, no. 1 (2005): 80–85. https://doi.org/10.5713/ajas.2006.80.

Cuppett, Susan L., and Clifford A. Hall. “Antioxidant Activity of the Labiatae”. Advances in Food and Nutrition Research 42 (1998): 245–71. https://doi.org/10.1016/s1043-4526(08)60097-2.

Ehrlinger, M. “Phytogenic Additives in Animal Nutrition.” Dissertation, Veterinary Faculty of the Ludwig Maximilians University, 2007.

Heinzl, I. “Efficient Microbiome Modulation with Phytomolecules”. EW Nutrition, August 30, 2022. https://ew-nutrition.com/pushing-microbiome-in-right-direction-phytomolecules/.

Hernández, F., J. Madrid, V. García, J. Orengo, and M.D. Megías. “Influence of Two Plant Extracts on Broilers Performance, Digestibility, and Digestive Organ Size.” Poultry Science 83, no. 2 (2004): 169–74. https://doi.org/10.1093/ps/83.2.169.

Kowalczyk, Adam, Martyna Przychodna, Sylwia Sopata, Agnieszka Bodalska, and Izabela Fecka. “Thymol and Thyme Essential Oil—New Insights into Selected Therapeutic Applications.” Molecules 25, no. 18 (2020): 4125. https://doi.org/10.3390/molecules25184125.

Lindner, , U. “Aromatic Plants – Cultivation and Use.” Düsseldorf: Teaching and Research Institute for Horticulture Auweiler-Friesdorf, 1987.

Oetting, Liliana Lotufo, Carlos Eduardo Utiyama, Pedro Agostinho Giani, Urbano dos Ruiz, and Valdomiro Shigueru Miyada. “Efeitos De Extratos Vegetais e Antimicrobianos Sobre a Digestibilidade Aparente, O Desempenho, a Morfometria Dos Órgãos e a Histologia Intestinal De Leitões Recém-Desmamados.” Revista Brasileira de Zootecnia 35, no. 4 (2006): 1389–97. https://doi.org/10.1590/s1516-35982006000500019.

Sadeh, Dganit, Nadav Nitzan, David Chaimovitsh, Alona Shachter, Murad Ghanim, and Nativ Dudai. “Interactive Effects of Genotype, Seasonality and Extraction Method on Chemical Compositions and Yield of Essential Oil from Rosemary (Rosmarinus Officinalis L”.).” Industrial Crops and Products 138 (2019): 111419. https://doi.org/10.1016/j.indcrop.2019.05.068.

Shabnum, Shazia, and Muzafar G. Wagay. “Essential Oil Composition of Thymus Vulgaris L. and Their Uses”. Journal of Research & Development 11 (2011): 83–94.

Sienkiewicz, Monika, Monika Łysakowska, Marta Pastuszka, Wojciech Bienias, and Edward Kowalczyk. “The Potential of Use Basil and Rosemary Essential Oils as Effective Antibacterial Agents.” Molecules 18, no. 8 (2013): 9334–51. https://doi.org/10.3390/molecules18089334.

Smith-Palmer, A., J. Stewart, and L. Fyfe. “Antimicrobial Properties of Plant Essential Oils and Essences against Five Important Food-Borne Pathogens”. Letters in Applied Microbiology 26, no. 2 (1998): 118–22. https://doi.org/10.1046/j.1472-765x.1998.00303.x.

Soković, Marina, Jelena Vukojević, Petar Marin, Dejan Brkić, Vlatka Vajs, and Leo Van Griensven. “Chemical Composition of Essential Oils of Thymus and Mentha Species and Their Antifungal Activities”. Molecules 14, no. 1 (2009): 238–49. https://doi.org/10.3390/molecules14010238.

Wei, Alfreda, and Takayuki Shibamoto. “Antioxidant Activities and Volatile Constituents of Various Essential Oils.” Journal of Agricultural and Food Chemistry 55, no. 5 (2007): 1737–42. https://doi.org/10.1021/jf062959x.

Yang, Li, Kui-Shan Wen, Xiao Ruan, Ying-Xian Zhao, Feng Wei, and Qiang Wang. “Response of Plant Secondary Metabolites to Environmental Factors”. Molecules 23, no. 4 (2018): 762. https://doi.org/10.3390/molecules23040762.

Özer, Hakan, Münevver Sökmen, Medine Güllüce, Ahmet Adigüzel, Fikrettin Şahin, Atalay Sökmen, Hamdullah Kiliç, and Özlem Bariş. “Chemical Composition and Antimicrobial and Antioxidant Activities of the Essential Oil and Methanol Extract of Hippomarathrum Microcarpum (Bieb.) from Turkey”. Journal of Agricultural and Food Chemistry 55, no. 3 (2007): 937–42. https://doi.org/10.1021/jf0624244.




Feed processing is the sustainability champion no one knows about (yet!)

Shutterstock

by Predrag Persak, Regional Technical Manager Europe, EW Nutrition

Imagine you’re at a pub quiz dedicated to feed production, and this question pops up: name a process that returns up to 25 times what was invested in it. Do you know the answer? I’m pretty sure you are probably using it every day: pelleting. For every unit of used energy, pelleting generates up to 25 times more in terms of the nutritional value for animals (mostly metabolizable energy).

The math is simple: while we gain 200 kcal/kg by pelleting broiler mash feed, only 10 Kilowatts are used to produce one ton of broiler feed. This is just one example of how sustainability is at the core of feed production – and has always been, long before it became a buzzword. So, to all those who operate feed mills, who take care of sourcing and quality, and to those behind numbers that represent nutritional values: You are pioneers of sustainability and should be proud of that.

How feed processing can drive sustainability efforts

Besides being proud, we must also be very responsible. Every nutritionist should focus on

  1. how processing of feed materials and feed influences the release of nutrients, nutrient density, and exclusion of antinutrients, and
  2. how processing can improve these dimensions, making feed more sustainable.

Do we take processing sufficiently into consideration? Do we create formulations in a dynamic or more static way? Not least in an era of precision feeding, the shift from static to dynamic is inevitable.

This is even clearer when we consider how processing can influence digestion, absorption, and the performance of animals. How so? Feed processing makes previously unusable materials suitable for nutrition or improves already usable materials. So, the feed processing itself is a key to sustainability.

 

Feed processing converts energy into more energy (?!)

Feed processing, in simple terms, means converting energy into more energy. This shouldn’t work, given the law of thermodynamics, but it does. Compound feed contains various feed materials and additives. Grains and protein sources (many times wrongly declared as byproducts), fibrous roughages and many other different components should not go together. Thanks to processing, they become feed which ensures the availability of all nutrients to the species, category, or animal production system for which that feed is intended.

Through processing, we alter the physical, chemical, and edible properties of used feed materials, making them usable for animals. Through proper processing, we improve the digestibility of feed materials by up to 20%, enabling a more effective – and thus more sustainable – use of feed resources. In practice, there is room for improvement to make feed processing even more of a sustainability champion.

Moisture optimization is key to energy-efficient pelleting

Let’s take a closer look at pelleting since it requires the most energy within feed processing. How much energy is used? This depends on many factors and can range from 5 KW/h up to 25. Pelleting is mostly used in broiler diets to reduce nutrient segregation and feed sorting and, by extension, feed wastage. Pelleting has also been found to increase the weight gain of individual birds and flock uniformity, and overall feed efficiency is higher.

Pelleting involves the agglomeration of mixed feed into whole pellets through a mechanical process using heat, moisture, and pressure (Falk, 1985). Heat (energy that is transported through steam) has the largest impact on pelleting efficacy. Steam injected during conditioning increases feed moisture and temperature, softens feed particles, extracts natural binders, and reduces friction which leads to greater production rates and pellet quality (Skoch et al., 1981).

The key to an efficient pelleting process is to set the parameters at the levels that will enable proper energy transfer from steam to feed particles. Besides steam quality, the moisture of the feed is a critical factor for efficient energy transfer. Generally, the thermal conductivity of the most used feed materials increases with increasing moisture. A level of 17% moisture in the conditioner is needed for efficient energy transfer. Below 17%, we need more steam (more energy) or more time (more capacity) to achieve the same result. That is why proper moisture optimization is needed to use the energy transferred through steam in the most efficient way.

Reduce shrinkage, improve sustainability

What about shrinkage? Shrinkage is not just a cost factor but a sustainability issue. We must not lose scarce and valuable materials and nutrients. Overall shrinkage tends to be around 1%. For global feed production as a whole, 1% annual shrinkage is equivalent to 15 years of Croatian compound feed production!

We help our industry to keep up sustainability efforts in terms of energy savings and shrinkage reduction by offering SurfAce. It’s a liquid preservative premixture with multiple economic and environmental benefits to the customer. It helps increase pellet output, improves conditioning, enhances the durability of the pelleted feed, reduces the formation of fines, and improves the overall quality of the final feed product. But most importantly, it optimizes feed production costs through energy savings and reduced labor input while also supporting the microbiological quality of the feed.

In the food sector, we have seen vast improvements in non-thermal food processing over the past decade. Examples include ultrasonication, cold plasma technology, supercritical technology, irradiation, pulsed electric field, high hydrostatic pressure, pulsed ultraviolet technology, and ozone treatment. I’m sure some of these technologies will be applied to feed processing one day. Until then, we must keep up our high sustainability standards and make it more efficient by applying all available tools in our feed processing toolbox.

References

Falk, D. “Pelleting Cost Center.” Essay. In Feed Manufacturing Technology III, edited by Robert R. McEllhiney, 167–90. Arlington, VA: American Feed Industry Association, 1981.

Skoch, E.R., K.C. Behnke, C.W. Deyoe, and S.F. Binder. “The Effect of Steam-Conditioning Rate on the Pelleting Process.” Animal Feed Science and Technology 6, no. 1 (1981): 83–90. https://doi.org/10.1016/0377-8401(81)90033-x.

 




Keep coccidiosis under control – naturally!

header image poultry broiler shutterstock 1733838041

By Inge Heinzl, Madalina Diaconu, Ajay Awati

Often you have an extensive coccidiosis control program in place. You don’t observe any clinical signs of coccidiosis. However, at the end of the cycle, you record significantly lower body weight and a higher FCR. There is a high probability that your animals have subclinical coccidiosis. This article digs deeper into understanding why birds don’t perform as they should, why subclinical coccidiosis occurs on the farm, and why drug resistance is an important factor.

Subclinical coccidiosis – a silent enemy

Clinical coccidiosis is clearly characterized by severe diarrhea, high mortality rates, reduced feed/water intake, and weight loss. By contrast, subclinical Coccidiosis does not display any visual signs and often remains undetected.

According to De Gussem (2008), the damages caused by subclinical coccidiosis can reach up to 70% of the total cost of coccidiosis control treatments, ranging from US$ 2.3 billion to US$ 13.8 billion/year in 2020 worldwide (De Gussem, 2008; Ferreira da Cunha, 2020; Blake et al., 2020).

Monitoring coccidiosis occurrence on the farm

There are several tools available to evaluate the level of infection. The most common ones are:

Lesion scoring – is used to evaluate the damages caused by coccidiosis in the intestinal tract. Lesion scoring gives insight into the severity of the infection. Furthermore, based on the location of lesions in the GI tract, it is possible to determine the plausible Eimeria spp. responsible for the infection.

OPG (Oocyst per gram) – the number of oocysts per gram of feces indicates the level of shedding of oocysts in the manure, litter, and, eventually, in the farm environment. OPG levels may not give the exact severity of the infection in the bird but certainly provide a clear idea of its likely spread within the flock.

Ways to deal with coccidiosis on the farm

Different tools are widely used to prevent and treat coccidiosis:

Anticoccidials:                  Chemicals, ionophores

Vaccination:                       Natural strains, attenuated strains

Bio-shuttle:                        Vaccine + ionophore

Natural anticoccidials:   Phytomolecules

These coccidiosis control programs are used depending on the farm history and the severity of the infection. Traditionally, treatment was heavily dependent on chemicals and ionophores. However, rampant and unbridled use of ionophores leads to resistance in Eimeria spp. on the farm, the failure of the control program, and significant performance losses, with high mortality due to coccidiosis. Therefore, the tools mentioned above are inserted in rotation or shuttle programs to minimize the generation of resistances. In a rotation program, the anticoccidial changes from flock to flock. In a shuttle program, the anticoccidial changes within one cycle according to the feed (Chapman, 1997).

However, this strategy is often not 100% effective due to a lack of diversity and overuse of certain tools within programs. The rigorous financial optimization of the program leads to the use of cost-effective but marginally effective solutions. These factors over the period weaken the program, which seems to work well but leads to resistance to anticoccidial drugs and sets up subclinical coccidiosis.

Resistances have been reported in the US (Jeffers, 1974, McDougald, 1981), South America (McDougald, 1987; Kawazoe and Di Fabio, 1994), Europe (Peeters et al., 1994; Bedrník et al., 1989; Stephan et al., 1997), Asia (Lan et al., 2017; Arabkhazaeli et al., 2013), and Africa (Ojimelukwe et al., 2018). Chapman and co-workers (1997) even stated that resistances were documented for all anticoccidial drugs employed at this time, and new products have not been approved for decades.

Resistance and subclinical coccidiosis can be approached naturally

When an anticoccidial has lost its effectiveness due to excessive use, some resistant coccidia survive. They can cause a mild course of the disease, subclinical coccidiosis, driving the costs high. Reducing the occurrence of resistance and subclinical coccidiosis can significantly decrease the expenses of coccidiosis control programs and, eventually, the cost of production.

Increasing consumer pressure to reduce the overall usage of drugs in animal production has driven innovation efforts to find natural solutions that can be effectively used within coccidiosis control programs. However, this shift was not easy for the producers. Lack of reliable data, poor understanding of the mode of action, lack of quality optimization, and unsubstantiated claims led to the failure of many earlier-generation natural solutions.

However, the consumer-driven movement to find natural solutions to animal gut health issues has recently led to relentless innovation in this area. Knowledge, research, and technological developments are now ready to offer solutions that can be an effective part of the coccidia control program and open opportunities to make poultry production even more sustainable by reducing drug dependency.

For centuries, phytomolecules have been used for their medicinal properties and effects on the health and well-being of animals and humans. In the case of coccidiosis, tannins and saponins have been proven to support animals in coping with this disease. Tannic acids and tannic acid extracts strengthen the intestinal barrier by reducing oxidative stress and inflammation (Tonda et al., 2018). On the other hand, saponins lessen the shedding of oocysts, improve the lesion score, and, in the case of an acute infection, the occurrence of bloody diarrhea (Youssef et al., 2021).

These natural substances can be integrated into shuttle or rotation programs to reduce the use of anticoccidials and, therefore, minimize resistance development.

Pretect D: Coccidiosis programs can be strengthened naturally!

In an EU field trial conducted with more than 200 000 birds, Pretect D (a natural phytogenic-based product designed to increase the efficacy of coccidiosis control) was used in the shuttle program together with ionophores. The trial provided excellent results on zootechnical performance (figures 1-4).

Figures 1-4: Zootechnical performance of broilers with Pretect D included in the shuttle program

Trials show that Pretect D supports the efficiency of coccidiosis control programs by impairing the Eimeria development cycle when used in combination with vaccines, ionophores, and chemicals as part of the shuttle or rotation program:

  • It protects the epithelium from inflammatory and oxidative damage
  • It promotes the restoration of the mucosal barrier function

Table 1 exemplifies one way of including a natural solution (Pretect D) in actual coccidiosis control programs.

Table 1: Exemple of including Pretect D into coccidiosis control programs

Natural solutions suit both farmers and consumers

With phytomolecules partly replacing anticoccidials in rotation or shuttle programs, the use of anticoccidials in poultry production can be decreased. On the one hand, this answers consumers’ demand; on the other hand, it leads to a push-back of resistances in the long run. The returning effectiveness of the anticoccidials can reduce subclinical coccidiosis, leading to lower costs spent on this disease and a higher profit for the farmers.

References:

Arabkhazaeli, F., M. Modrisanei, S. Nabian, B. Mansoori, and A. Madani. “Evaluating the Resistance of Eimeria spp. Field Isolates to Anticoccidial Drugs Using Three Different Indices.” Iran J Parasitol. 8, no. 2 (2013): 234–41.

Bedrník, P., P. Jurkovič, J. Kučera, and A. Firmanová. “Cross Resistance to the IONOPHOROUS Polyether Anticoccidial Drugs IN Eimeria Tenella Isolates from Czechoslovakia.” Poultry Science 68, no. 1 (1989): 89–93. https://doi.org/10.3382/ps.0680089

Blake, Damer P., Jolene Knox, Ben Dehaeck, Ben Huntington, Thilak Rathinam, Venu Ravipati, Simeon Ayoade, et al. “Re-Calculating the Cost of Coccidiosis in Chickens.” Veterinary Research 51, no. 1 (2020). https://doi.org/10.1186/s13567-020-00837-2

Chapman, H. D. “Biochemical, Genetic and Applied Aspects of Drug Resistance in Eimeria Parasites of the Fowl.” Avian Pathology 26, no. 2 (1997): 221–44. https://doi.org/10.1080/03079459708419208.

De Gussem, M., and S. Huang. “The Control of Coccidiosis in Poultry.” International Poultry Production 16, no. 5 (2008): 7–9.

Ferreira da Cunha, Anderson, Elizabeth Santin, and Michael Kogut. “Editorial: Poultry Coccidiosis: Strategies to Understand and Control.” Frontiers in Veterinary Science 7 (2020). https://doi.org/10.3389/fvets.2020.599322

Jeffers, T. K. “Eimeria Acervulina and E. Maxima: Incidence and Anticoccidial Drug Resistance of Isolants in Major Broiler-Producing Areas.” Avian Diseases 18, no. 3 (1974): 331. https://doi.org/10.2307/1589101

Kawazoe, Urara, and J. Di Fabio. “Resistance to DICLAZURIL in Field Isolates OfEimeriaspecies Obtained from Commercial BROILER Flocks in Brazil.” Avian Pathology 23, no. 2 (1994): 305–11. https://doi.org/10.1080/03079459408418998

Lan, L.-H., B.-B. Sun, B.-X.-Z. Zuo, X.-Q. Chen, and A.-F. Du. “Prevalence and Drug Resistance of Avian Eimeria Species in Broiler Chicken Farms of Zhejiang PROVINCE, CHINA.” Poultry Science 96, no. 7 (2017): 2104–9. https://doi.org/10.3382/ps/pew499

McDougald, L. R. “Anticoccidial Drug Resistance in the Southeastern United STATES: POLYETHER, IONOPHOROUS Drugs.” Avian Diseases 25, no. 3 (1981): 600. https://doi.org/10.2307/1589990

McDougald, Larry R., Jose Maria Silva, Juan Solis, and Mauricio Braga. “A Survey of Sensitivity to Anticoccidial Drugs in 60 Isolates of Coccidia from Broiler Chickens in Brazil and Argentina.” Avian Diseases 31, no. 2 (1987): 287. https://doi.org/10.2307/1590874

Ojimelukwe, Agatha E., Deborah E. Emedhem, Gabriel O. Agu, Florence O. Nduka, and Austin E. Abah. “Populations of Eimeria Tenella Express Resistance to Commonly Used Anticoccidial Drugs in Southern Nigeria.” International Journal of Veterinary Science and Medicine 6, no. 2 (2018): 192–200. https://doi.org/10.1016/j.ijvsm.2018.06.003

Peeters, Johan E., Jef Derijcke, Mark Verlinden, and Ria Wyffels. “Sensitivity of AVIAN EIMERIA Spp. to Seven Chemical and Five Ionophore Anticoccidials in Five Belgian INTEGRATED Broiler Operations.” Avian Diseases 38, no. 3 (1994): 483. https://doi.org/10.2307/1592069

Stephan, B., M. Rommel, A. Daugschies, and A. Haberkorn. “Studies of Resistance to Anticoccidials IN Eimeria Field Isolates and Pure Eimeria Strains.” Veterinary Parasitology 69, no. 1-2 (1997): 19–29. https://doi.org/10.1016/s0304-4017(96)01096-5

Tonda, RM, J.K. Rubach, B.S. Lumpkins, G.F. Mathis, and M.J. Poss. “Effects of Tannic Acid Extract on Performance and Intestinal Health of Broiler Chickens Following Coccidiosis Vaccination and/or a Mixed-Species Eimeria Challenge.” Poultry Science 97, no. 9 (2018): 3031–42. https://doi.org/10.3382/ps/pey158

Youssef, Ibrahim M., Klaus Männer, and Jürgen Zentek. “Effect of Essential Oils or Saponins Alone or in Combination on Productive Performance, Intestinal Morphology and Digestive Enzymes’ Activity of Broiler Chickens.” Journal of Animal Physiology and Animal Nutrition 105, no. 1 (2020): 99–107. https://doi.org/10.1111/jpn.13431




How to mitigate quality degradation in broiler breasts

white chickens farm

By Inge Heinzl and Ajay Bhoyar, EW Nutrition

Faster growth of breast muscle in broilers may lead to increased incidences of different types of muscle degeneration. Downgrading the affected breast fillets results in high economic losses for the poultry meat industry.

The article discusses the three important myopathies impairing the breast muscles, their impact on the meat industry, influencing factors, and how to cope with these challenges.

Muscle degeneration heaps up with faster broiler growth

According to Sirri and co-workers (2016), breast fillets from broilers with 3.9 kg live weight carry a higher risk for myopathic lesions. Studies in different countries revealed that myopathies in broilers are not neglectable:

Country Myopathy Number of breasts examined Conditions Occurrence Reference
Italy WS 28,000 broilers commercial 12 % Petracci et al., 2013
Italy WS 70 flocks; always 500 of 35,000 breasts randomly examined commercial 43%, with 6.2% considered severe Lorenzi et al., 2014
Italy WS 57 flocks commercial 70.2 % (medium)-82.5 % (heavy-weight) Russo et al., 2015
Italy WS 16,000 samples commercial 9 % moderate22 % severe Petracci in Baldi et al., 2020
Brazil WS 25,520 commercial 10 % Ferreira et al., 2014
USA WS 960 (week 6)+ 960 (week 9) experimental Score 1: 78.4 % (wk 6)
29.9 % (wk 9)
Score 2: 14.0 % (wk 6)
53.9 % (wk 9)
Score 3:0 % (wk 6)
15.1 % (wk 9)
Kuttapan et al., 2017
Brazil WB commercial 10-20 % Carvalho, in Petracci et al., 2019
Italy WB 16,000 samples commercial 42 % moderate
18 % severe
Petracci, in Baldi et al., 2020
China WB 1,135 breast fillets commercial 61.9% Xing et al., 2020
USA WB 960 (week 6)+ 960 (week 9) experimental Score 1: 32.5 % (wk 6)
33.2 % (wk 9)
Score 2: 7.9 % (wk 6)
36 % (wk 9)
Score 3: 1.96 % (wk 6)
15.6 % (wk 9)
Kuttapan et al., 2017
Italy SM 16,000 samples commercial 4 % moderate
17 % severe
Petracci in Baldi et al., 2020
Brazil SM 5,580 samples commercial 10 % Montagna et al., 2019

 

Figure 1: Different myopathies in broilers (R. Baileys)

As the appearance of products is one of the most important arguments for the purchase decision, these myopathies are serious issues; the downgrading of the breast quality results in a lower reward for the producer. Kuttapan et al. (2016) estimated that 90 % of the broilers are affected by wooden breast and white striping (see below), causing about $200 million to $1 billion of economic losses to the U.S. poultry industry per year.

Wooden Breast (WB), a result of the proliferation of connective tissues

The muscle affected by the wooden breast is bulging and hard, is covered with clear, viscous fluid, and shows petechiae (see figure 2). The myopathy of the pectoralis major is “pale expansive areas of substantial hardness accompanied by white striation” (Kuttapan, 2016; Huang and Ahn, 2018; Sihvo et al., 2013). It is characterized by microscopically visible polyphasic myodegenerations with fibrosis in the chronic phase. At approximately two weeks of age, it appears as a focal lesion but then develops as a widespread fibrotic injury (Papah et al., 2017). WB can be detected by palpating the breast of the live bird.

Figure 2: Comparison of a severe wooden breast (on the left) and a healthy breast fillet (on the right)

Source: Kuttapan et al., 2016

According to Kuttapan et al. (2016), the anomaly is caused by circulatory insufficiency and increased oxidative stress resulting in damage and degeneration. Its occurrence rose with increasing growth and slaughter weights of the birds. Wooden breast is more common in male than female broilers as they show an increased expression of genes related to the proliferation of connective tissues (Baldi et al., 2021).

The hardness of the meat, a 1.2 – 1.3 % higher fat content (Soglia et al., 2016, Tasoniero et al., 2016), and the worse appearance lead to a degradation of the fillet quality (Kuttappan et al., 2012). The reduction in the water holding capacity of muscle results in toughness before and after cooking.

White Striping (WS), a result of fiber degeneration

The characteristics of WS are white striations parallel to the muscle fibers. A microscopic examination of these white stripes reveals an accumulation of lipids and a proliferation of connective tissue occurring in breast fillets and thighs (Kuttappan et al., 2013a; Huang and Ahn, 2018). Kuttapan et al. (2016) adapted a scoring system for the evaluation of the severity of WS, which he had established earlier (Kuttapan et al., 2012)(see picture 1). It was concluded that broilers fed a diet with high energy content led to higher and more efficient growth (improved feed conversion, higher live and fillet weights) but also to a higher percentage of fillets showing a severe degree of white striping.

Figure 3: Different degrees of white striping

  • 0 = normal (no distinct white lines)
  • 1 = moderate (small white lines, generally < 1 mm thick)
  • 2 = severe (large white lines, 1-2 mm thick, very visible on the fillet surface)
  • 3 = extreme (thick white bands, > 2 mm thickness, covering almost the entire surface of the fillet
  • (scoring and image source: Kuttapan, 2016)

 

Moreover, the WB and WS can simultaneously occur in the same muscle (Cruz et al., 2016; Kuttappan, Hargis, & Owens, 2016; Livingston, Landon, Barnes, & Brake, 2018).

Spaghetti Meat (SM), a result of decreased collagen linking

The condition of Spaghetti Meat was first mentioned by Bilgili (2015) under “Stringy-spongy”. SM is characterized by an insufficient bonding of the muscles due to an immature intramuscular connective tissue in the pectoralis major. The fiber bundles composing the breast muscle detach, and the muscle gets soft and mushy and resembles spaghetti pasta (Baldi et al., 2021). Probably due to the reduced collagen-linking degree, the texture of SM fillets is smoother after cooking (Baldi et al., 2019). In contrast to wooden breast, SM cannot be noticed in the living animal. Meat severely impacted by SM is downgraded and can only be used in further processed products, whereas slightly affected meat can be sold in fresh retailing (Petracci et al., 2019).

Another possible explanation for this myopathy may be the strong development of the breast muscle. The thickness of its upper section might reduce muscular oxygenation by compressing the pectoral artery (Soglia et al., 2021). The spaghetti structure generally appears mainly in the superficial layer and less in the deep ones.

Oxidative stress – one link in the chain of causes for myopathies

Oxidative stress is a result of impaired blood supply

Oxidative stress is one key factor of myopathies in breast muscle. As the faster growth is connected with an increase in muscle fiber diameter, the higher pressure of the surrounding fascia on the muscle tissue compresses the blood vessels, leading to a decreased blood flow, resulting in insufficient oxygen supply (hypoxia) and limited removal of metabolic by-products (Lilburn et al., 2019) from the muscle tissue. Hypoxia as – well as hyperoxia – plus the deficient removal of metabolic waste, promote the generation of free radicals (Kähler et al., 2016; Strapazzon et al., 2016; Petrazzi et al., 2019). If the endogenous antioxidant system cannot efficiently eliminate these ROS by using endogenous and exogenous antioxidants, the ultimate effect is increased oxidative stress.

Soglia and co-workers (2016) reported higher TBARS (Thiobarbituric acid reactive substances) and protein carbonyl levels, signs of oxidative stress, in severe wooden breast muscle tissue. The oxidative stress hypothesis was also supported by gene transcription analysis conducted by Mutryn et al. (2015) and Zambonelli et al. (2017).

Oxidative stress causes damage

ROS (reactive oxygen species) or free radicals are highly reactive. They can cause damage to the DNA, RNA, proteins, and lipids in the muscle cells (Surai et al., 2015), leading to inflammation and metabolic disturbances, and, in the end, the degeneration of muscle fibers (Kuttapan et al., 2021). If the regenerative capacity of the muscle cells does not countervail against the damages caused by oxidative stress, fibrous tissue and fat accumulate and lead to myopathies such as wooden breast (Petracci et al., 2019)

Oxidative stress can be managed

To support the animals in coping with oxidative stress, combining two approaches, an external and an internal, makes sense. This entails protecting feed at the same time as protecting the animal.

Chemical antioxidants preserve feed quality and prevent oxidation

Chemical antioxidants such as ethoxyquin, BHA, and BHT efficiently prevent feed oxidation. These antioxidants prevent the oxidation of unsaturated fats/oils and maintain their energy value. They are scavengers for free radicals, protect trace minerals like Zn, Cu, Mg, Se, and Vit E from oxidation and spare them to be used in the body for different metabolic processes as well as for the endogenous antioxidant system.

However, keep in mind that chemical antioxidants are strictly regulated, depending on type, concentration, and region. Ethoxyquin has a challenging status in the EU, for instance, due to a ruling that excludes it for the use of long-living or reproductive animals and that sets safety levels at a maximum total concentration of 50 mg ethoxyquin/kg complete feed for all animal species, except dairy ruminants.

Phy­tomolecules act as natural antioxidants and reduce lipid oxidation in breast muscles

Inside the body, phy­tomolecules help to mitigate oxidative stress by the direct scavenging of ROS and the activation of antioxidant enzymes. Phytogenic compounds like Carvacrol and thymol possess phenolic OH-groups that act as hydrogen donors (Yanishlieva et al., 1999). These hydrogens can “neutralize” the peroxy radicals produced during the first step of lipid oxidation and, therefore, retard the hydroxyl peroxide formation. The increase in serum antioxidant enzyme activities and a resulting lower level of malondialdehyde (MDA) can be caused by cinnamaldehyde (Lin et al., 2003). MDA is a highly reactive dialdehyde generated as a metabolite in the degradation process of polyunsaturated fatty acids.

Antioxidant capacity of phytomolecules demonstrated in broilers

A trial with 480 Cobb male chicks (3 treatments, 8 replicates) was conducted at the University of Viçosa (Brazil). The breast muscles of the birds fed a blend of phy­tomolecules showed lower MDA levels and thus reduced lipid oxidation compared to the negative control, but also to the birds fed an antibiotic.

The impact of breast muscle degradation in broilers can be mitigated

The downgrading of broiler meat due to increased incidence of breast muscle myopathies is a common issue, resulting in the significant economic losses to the broiler meat producers. Oxidative stress caused due to due faster growth rate and various other stressors, including the oxidation of feed and feed ingredients, can contribute to increased incidence of woody breast and white striping. Different nutritional and management strategies are employed to reduce WB and WS in broiler production. The inclusion of synthetic antioxidants to control the oxidation in feed as well as phytomolecules to support the endogenous antioxidant system can be a part of promising tools to mitigate the impact of breast myopathies and reduce economic losses in broiler production.

 

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