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Meat quality is a result of genetics, feeding, the microbiome, and the handling of animals and meat

Different Pieces Of Meat Shutterstock

by Dr. Inge Heinzl, Editor EW Nutrition

Nowadays, nutrition is no longer about pure nutrient intake; enjoyment is also a priority. Consumers attach great importance to the high quality of food and, therefore, also of meat. The genetic selection for faster growth and feeding high-energy diets made meat production more efficient and shortened the raising period. However, this selection may sometimes also result in challenges to meat quality, such as worse water holding capacity, less marbling, less flavor, and reduced storage & processing properties.

The following article will provide detailed information about what meat quality is, how the gut microbiota influences it, and how we can increase meat quality by feeding and modulating the intestinal microflora.

Which factors can contribute to meat quality?

Meat quality is a complex term. On the one hand, meat quality covers measurable parameters such as the content of nutrients, moisture, microbial contamination, etc. On the other hand, and to no small extent, the consumers’ preferences are significant. Since meat today is often sold as cuts or in parts (e.g., broiler drumsticks, breast), processing also affects the quality of meat and meat products.

Physical characteristics are objective determinants of meat quality

Physical characteristics are parameters that can be measured. For meat, the following measurable parameters determine meat quality:

1.  Fat content and fatty acid composition influence tenderness and taste

Some years ago, the majority of consumers asked for completely lean meat, which, fortunately, has now changed. Fat is a flavor carrier. Especially intramuscular fat (marbling) melts during the preparation, making the meat tender, juicy, and taste good. Fat also transports fat-soluble vitamins.

A further criterion is the composition of the fat, the fatty acids. Geese fat, e.g., is known for its high content of oleic, linoleic, linolenic, and arachidonic acid, all of them derivates of the enzymatic denaturation of stearic acid (Okruszek, 2012).

One exception is cholesterol. Although belonging to the lipids and improving the sensory quality of meat, consumers prefer meat with low cholesterol content.

2.  Protein and amino acid content influence the meat value

The content and the composition of protein are important factors in meat quality. Protein is essential for constructing and maintaining organs and muscles and for the functionality of enzymes. The human body needs 20 different amino acids for these tasks, eleven of which it can manufacture by itself. Nine amino acids, however, must be provided by food and are called essential amino acids. Meat is a highly valuable protein source, rich in protein and essential amino acids. The protein quality, therefore, includes the chemical and amino acid score, the index for essential amino acids, and the biological value.

In addition to the pure nutritional value, amino acids contribute to flavor and taste. These flavor amino acids directly influence meat’s freshness and flavor and include threonine, alanine, serine, lysine, proline, hydroxyproline, glutamic acid (glutamate is important for the umami taste), aspartic acid, and arginine.

3.  Vitamins and trace elements are essential nutrients

Meat is a primary source of B vitamins (B1-B9) and, together with other animal products such as eggs and milk, the only provider of Vitamin B12. Vitamin A is available in the innards, vitamin D in the liver and fat fish, and vitamin K in the flesh.

The most important mineral compounds in meat are zinc, selenium, and iron. Humans can utilize the iron from animal sources particularly well.

4.  pH and speed of pH decline decide if the meat is suited for cooking

Since broiler chicken meat nowadays is usually consumed as cut-up pieces or processed products, the appearance at the meat counter or in the plastic box is essential for being sold. The color, seen as an apparent measurement of the freshness and quality of the meat, is influenced by the pH. The muscle pH post-mortem plays an essential role in meat quality. Due to the glycolytic process, the pH post-mortem is a good indication for evaluating physiological meat quality. A rapid pH decline post-mortem to 5.8-6.0 in most cases leads to pale, soft, and exudative (PSE) meat with reduced water retention (Džinić et al., 2015), whereas a high ultimate pH results in dark, firm, and dry (DFD) meat with poor storage quality (Allen et al., 1997)

5.  Nobody wants meat like leather

The shear force is a measure of the tenderness of the meat. To determine the shear force, the meat undergoes the process of cooking and chilling. Afterward, standardized meat blocks, with fibers running along the length of the sample, are put into the Warner-Bratzler system. The blade used simulates teeth, and the system measures the force necessary to tear the piece of meat.

6.  Microbial contamination is a no-go

The microbial contamination of the meat often occurs during the slaughter process. Let’s take a look at salmonella or campylobacter in poultry. The chickens take up salmonella with contaminated feed or water. Campylobacter is transmitted by infected wild birds, inadequately cleaned and disinfected cages, or contaminated water. The bacteria proliferate in the intestine. At slaughter, the intestine’s microorganisms can spread onto the meat intended for human consumption.

7.  High water holding capacity is necessary to have tender meat

The moisture content contributes to the meat’s juiciness and tenderness and improves its quality. If the meat loses its moisture, it gets tough, and quality decreases. Additionally, drip loss reduces the nutritional value of meat and its flavor.

8.  Fat oxidation makes meat rancid, and oxidative stress can cause myopathies in broiler breasts

Rancidity of meat occurs when the fat in the flesh gets oxidized. There are different signs of meat rancidity: bad odor, changed color, and a sticky, slimy texture. Poultry meat is considered more susceptible to the development of oxidative rancidity than red meat. This can be explained by its higher content of phospholipids, PUFAs, especially in the thighs. The breast meat, however, has a relatively low level of intramuscular fat (up to 2 %) and, additionally, myoglobin is a natural antioxidant.

But oxidative stress in broiler breasts – and this more and more happens due to a selection of always bigger breasts – can lead to muscle myopathies such as white stripes or wooden breasts, making the meat only usable for processed products.

Sensory meat quality addresses the human senses

Besides physical quality, the sensory and chemical characteristics are essential to meat’s economic importance. All attributes of meat that stimulate the human senses (vision, smell, taste, and touch) belong to the sensory quality. It, therefore, is more subjective and hard to determine. The most important features for the consumer include color (attractive or unattractive), texture (tenderness, juiciness, marbling, drip loss), and taste/ flavor (Thorslund et al., 2016).

The appearance is the first impression

Nowadays, meat is often sold as cuts lying in polystyrene or clear plastic trays, over-wrapped with transparent plastic films, so the appearance is paramount. The meat must show an attractive color. Muscle myopathies, such as the ones occurring in chickens, would not meet consumers’ needs.

How does the flavor of meat develop?

There is a reaction between reducing sugars and amino acids when meat is cooked (Mottram, 1998). This Maillard reaction, along with the degradation of vitamins, lipid oxidation, and their interaction, is responsible for the production of the volatile flavor components forming the characteristic aroma and flavor of cooked meat (MacLeod, 1994). Werkhoff et al. (1990) consider cysteine and methionine the most significant contributors to meat flavor development. One factor deteriorating this quality characteristic is lipid peroxidation, which turns the taste to rancid.

Some sensory characteristics are related to physical ones

The parameters of sensory meat quality can be partly explained by measurable parameters. Water retention, e.g., influences the juiciness of the meat. The palatability increases with higher intramuscular fat or marbling (Stewart et al., 2021), the initial pH and the speed of decline decide if the flesh will be pale, soft, and exudative or normal, and lipid peroxidation is the leading cause of a decrease in meat quality (Pereira & Abreu, 2018).

Processing quality

For the processing quality, muscle structure, chemical ingredient interactions, and muscle post-mortem changes are decisive (Berri, 2000).

Does the microbiome influence the meat quality?

The gastrointestinal tract of monogastric animals disposes of a microbiome of primarily bacteria, mainly anaerobic Gram-positive ones (Richards et al., 2005). With its complex microbial community, the digestive tract is responsible for digesting feed and absorbing nutrients, but also for eliminating pathogens and developing immunity. Gut microbiotas play an essential role in digestion, are decisive concerning the synthesis of fatty acids, proteins, and vitamins, and, therefore, influence meat quality (Chen, 2022).

Intestinal microbiotas vary by species/breeds and age (Ma et al., 2022; Sun et al., 2018), and so does meat quality. For example, Duroc pigs with meat of high tenderness, good flavor, and excellent tastiness show different microbiota than other breeds (Xiao, 2017). Zhao et al.(2022) examined high- and low-fat Jinhua pigs, with the high-fat pigs showing more increased backfat thickness but also a higher fat content in the longissimus dorsi. They found low-fat pigs showed a higher abundance of Prevotella and Bacteroides, Ruminococcus sp. AF12-5, Faecalibacterium sp.OFO4-11AC und Oscillibacter sp. CAG:155, which are all involved in fiber fermentation and butyrate production. The high-fat animals showed a higher abundance of Firmicutes and Tenericutes, indicating that they are responsible for higher fat production of the organism in general but also a better fat disposition in the flesh. Lei et al. (2022) showed that abdominal fat was positively correlated with the occurrence of Lachnochlostridium and Christensenelleceae.

The intestinal microbiota-muscle axis enables us to improve meat quality by controlling intestinal microbiota (Lei, 2022). However, to develop strategies to enhance the quality of meat, understanding the composition of the microbiota, the functions of the key bacteria, and the interaction between the host and microbiota is of utmost importance (Chen et al., 2022).

Different factors influence the microbiome

Apart from that microbiotas are different in different breeds, they are additionally influenced by diseases, feeding (diets, medical treatments with, e.g., antibiotics), and the environment (climate, geographical position). This could be shown by different trials. The genetic influence on microbiota was impressively documented by Goodrich et al. (2014), who detected that the microbiomes of monozygotic twins differ less than the ones of dizygotic twins. Lei et al. (2022) compared the microbiota of two broiler breeds (Arbor Acres and Beijing-You, the last one with a higher abdominal fat rate) and found remarkable differences in their microbiota composition. When raising them in the same environment and with the same feed, the microbiotas became similar. Zhou et al. (2016) contrasted the cecal microbiota of five Tibetan chickens from five different geographic regions with Lohmann egg-laying hens and Daheng broiler chickens. Besides seeing a difference between the breeds, slightly distinct microbiota between the regions could also be noticed.

The intestinal microbiome can actively be changed by

  • promoting the wanted microbes by feeding the appropriate nutrients (e.g., prebiotics)
  • reducing the harmful ones by fighting them, for example, with organic acids or phytomolecules
  • directly applying probiotics and adding, therefore, desired microbes to the microbiome.

An increase in the abundance of Lactobacillus and Succiniclasticum could be achieved in pigs by feeding them a fermented diet, and Mitsuokella and Erysipelotrichaceae proliferated by adding a probiotic containing B. subtilis and E. faecalis to the diet (Wang et al., 2022).

How to change the intestinal microbiome to improve meat quality?

Before changing the microbiome, we must know which microbes are “responsible” for which characteristics. However, the microbiotas do not act individually but as consortia. The following table shows a selection of bacteria that, besides supporting the gut and its functions, influence meat quality in some way.

Metabolites Producing bacteria Biological functions and effects on pigs
Short-chain fatty acids (acetate, butyrate, and propionate) Ruminococcaceae

Ruminococcus

Lachnospiraceae

Blautia

Roseburia

Lactobacillaceae

Clostridium

Eubacterium

Faecalibacterium

Bifidobacterium

Bacteroides

Regulate lipid metabolism

Improve meat quality

Lactate Lactic acid bacteria

Bifidobacterium

Important metabolite for cross-feeding of SCFA-producing microbiota
Bile acids (primary and secondary bile acids) Clostridium species

Eubacterium

Parabacteroides

Lachnospiraceae

Regulate lipid metabolism
Ammonia Amino acid fermenting commensals

Helicobacter

By-product of amino acid fermentation

Inhibits short-chain fatty acid oxidation

B Vitamins and vitamin K Bacteroides

Lactobacillus

Serve as coenzymes in neurological processes (B vitamins)

• Essential vitamin for proper blood clotting (vitamin K)

Table 1: Bacteria influencing meat quality (according to Vasquez et al., 2022)

Fat for meat quality is intramuscular fat

If we talk about increasing fat to improve meat quality, we talk about increasing intramuscular fat or marbling, not depot fat. The fat in meat-producing animals is mostly a combination of triglycerides from the diet and fatty acids synthesized. Fat deposition and composition in non-ruminants reflect the fatty acid composition of the diet but are also closely related to the design of the microbiome; short-chain fatty acids in monogastric, e.g., are exclusively produced by the gut microbiome (Dinh et al., 2021; Vasquez et al., 2022). Intramuscular fat is mainly made of triglycerides but also disposes of phospholipids associated with proteins, such as lipoproteins or proteolipids, influencing meat flavor. The fermentation of indigestible polysaccharides or amino acids results in short-chain or branched-chain fatty acids, respectively. Lactate, produced by lactic acid bacteria, is utilized by SCFA-producing microbiota. An imbalance in the microbiome fosters lipid deposition, as shown by Kallus and Brandt (2012), who found a higher proportion of Firmicutes to Bacteroidetes (50% higher) in obese mice than in lean ones. In a trial described by Zhou et al. (2016), tiny Tibetian chickens with a low percentage of abdominal fat were compared to two breeds (Lohmann layers and Daheng broilers) being large and with a high percentage of abdominal fat. The Tibetan chickens showed a two to four-fold higher abundance of Christensenellacea in the cecal microbiome. Christensenellas belong to the bacterial strain of firmicutes. They are linked to slimness in human nutrition, which was already proven by Goodrich et al. (2014) and is the contrary stated by Lei et al. (2022).

Another example was provided by Wen et al. (2023). They compared two broiler enterotypes distinguished by Clostridia vadinB60 and Rikenellaceae_RC9_gut and saw that the type with an abundance of Clostridia_vadinBB60 showed higher intramuscular fat content but also more subcutaneous fat tissue. The scientists also found another bacterium especially responsible for intramuscular fat: A lower plethora of Clostridia vadimBE97 resulted in a higher intramuscular fat content in breast and thigh muscles but not adipose tissues. Similar results were achieved in a trial with pigs and mice: Jinhua pigs showed a significantly higher level of intramuscular fat than Landrace pigs. When transplanting the fecal microbiota of the two breeds in mice, the mice showed similar characteristics in fat metabolism as their donors of feces (Wu et al., 2021).

According to several studies (e.g., Chen et al., 2008; Liu et al., 2019), intramuscular fat in chicken has a low heritability but may be controlled by feeding up to a certain extent. In pigs, Lo et al. (1992) and Ding et al. (2019) found a moderate to low (0.16 – 0.23) heritability for intramuscular fat, but Cabling et al. (2015) calculated a heritability of 0.79 for the marbling score.

At least, especially the composition of fatty acids can easily be changed in monogastric (Aaslyng and Meinert, 2017). Zou et al. (2017) examined the effect of Lactobacillus brevis and tea polyphenol, each alone or combining both. Lactobacillus is probably involved in turning complex carbohydrates into metabolites lactose and ethanol, but also acetic acid and SCFA. SCFAs are mainly produced by Saccharolytic and anaerobic microbiota, aiding in the degradation of carbohydrates the host cannot digest (e.g., cellulose or resistant polysaccharides into monomeric and dimeric sugars and fermenting them subsequently into short-chain fatty acids). Including fibers and various oligosaccharides was shown to increase the gut microbiome’s fermentation capacity for producing short-chain fatty acids.

In a trial conducted by Jiao et al. (2020), they showed that SCFAs applied in the ileum modulate lipid metabolism and lead to higher meat quality in growing pigs. A plant polyphenol was used by Yu et al. (2021). The added resveratrol, a plant polyphenol in grapes and grape products, to the diet of Peking ducks and could significantly increase intramuscular fat.

Oxidation of lipids and proteins must be prevented

The composition of the fatty acids and occurring oxidative stress in adipose and muscle tissue influences or impacts meat quality in farm animals (Chen et al., 2022). During the last few years, the demand for healthier animal products containing higher levels of polyunsaturated fatty acids has increased. Consequently, the risk of lipoperoxidation has risen (Serra et al., 2021). Solutions are needed to counteract this deterioration of meat quality. As can be seen in table 1, ammonia produced by amino acid-fermenting commensals and Helicobacter inhibits the oxidation of SCFAs. Ma et al. (2022) changed the microbiome of sows by feeding a probiotic from mating till day 21 of lactation and achieved a decreased level of MDA, a sign of reduced oxidative stress. Similar results were achieved by He et al. (2022). In their trial, the supplementation of 200 mg yeast ß-glucan/kg of feed significantly decreased the abundance of the phylum WPS-2 as well as markedly increased catalase, superoxide dismutase (both p<0.05) and the total antioxidant activity (p<0.01) in skeletal muscle. Another approach was done by Wu et al. (2020) in broilers. They applied glucose oxidases (GOD) produced by Aspergillus niger and Penicillium amagasakiense. Both enzymes did not disturb but improved beneficial bacteria and microbiota. The GOD produced by A. niger reduced the content of malondialdehyde in the plasma.

Another alternative is antioxidant extracts from plants (Džinić, 2015). As consumers nowadays bet more on natural products, they would be good candidates. They are considered safe and, therefore, well-accepted by consumers and have beneficial effects on animal health, welfare, and production performance.

Hazrati et al. (2020) showed in a trial that the essential oils of ajwain and dill decreased the concentration of malondialdehyde (MDA) in quails’ breast meat and, therefore, lipid peroxidation and reduced cooking loss. The antioxidant effects of thymol and carvacrol were shown by Luna et al. (2010). The group receiving the essential oils showed lower TBARS in the thigh samples than the control group but similar TBARS to the butylated hydroxytoluene-provided group.

Protein quality is a question of essential amino acids

Protein with a high content of essential amino acids is one of the most critical components of meat. Alfaig et al. (2014) tested probiotics and thyme essential oil in broilers. They found out that the content of EAAs in breast and thigh muscles numerically increased gradually from the control over the probiotic and a combination of a probiotic up to the thyme essential oil group. A significant (p<0.05) increase in all tested amino acids (arginine, cysteine, phenylalanine, histidine, isoleucine, leucine, lysine, methionine, threonine, and valine) could be observed in the samples of the breast and the thigh muscles when comparing the thyme essential oil group with the control. Zou et al. (2017) provided similar results, showing a significant increase in leucine and glutamic acid as well as a numerical increase in lysin, valine, methionine, isoleucine, phenylalanine, threonine, asparagine, alanine, glycin, serin, and proline through the addition of a combination of Lactobacillus brevis and tea polyphenols. They also determined an increase in the beneficial bacteria Lactobacillus and Bacteroides. The experimental results led them to the assumption that both additives may also improve the taste of meat by increasing some of the essential and delicate flavors produced by amino acids.

Tenderness is closely related to drip loss

The already mentioned trial conducted by Lei et al. (2022) with two different broiler breeds (Arbor Acres and Beijing-You) having different microbiota showed a negative correlation between drip loss and the abundance of Lachnochlostridium. They remodeled the Arbor Acres’ microbiome by applying a bacterial suspension derived from the Beijing-You breed and decreased drip loss in their meat. He et al. (2022) changed the microbiome by adding yeast ß-glucan to the diet of finisher pigs. They achieved a reduced cooking loss (linear, p<0.05) and a lower drip loss (p<0.05), together indicating a better water-holding capacity, as well as a decreased lactate content. The addition of a multi-species probiotic to the diet of finishing pigs tended to result in lower cooking and drip loss(p<0.1) besides modulating the intestinal flora (higher lactobacilli and lower E. coli counts in the feces) (Balasubramanian et al., 2017) and the inclusion of Lactobacillus brevis and tea polyphenol individually or in a synergistic combination improved water holding capacity and decreased drip loss Zou et al. (2017).

Puvača et al. (2019) observed the lowest drip-loss values in breast meat and thigh with drumstick through feeding chickens 0.5 g or 1.0 g of hot red pepper per 100 g of feed, respectively, in the grower and finisher phase. The feeding of resveratrol reduced drip loss of Peking ducks’ leg muscles. SCFA infused into the ileum enlarged the longissimus dorsi area and alleviated drip loss (Jiao et al, 2021).

The decrease and increase of the pH after slaughtering determines meat quality

The pH in the muscles of a living animal is about 7.2. With slaughtering and bleeding, the energy supply of the muscles is interrupted. The stored glycogen gets degraded to lactic acid, lowering the pH. Usually, the lowest pH value of 5.4-5.7 in meat is reached after 18 to 24 hours. Afterward, it starts to rise again.

In stressed animals, the stress hormones adrenalin and noradrenalin provoke a rushly occurring and, due to a lack of oxygen, anaerobic metabolism and the quick production of lactic acid. This too rapid decrease in pH leads to the denaturation of proteins in the muscle cells and reduced water-holding capacity. The result is PSE (pale, soft, and exudative) meat.

On the contrary, DFD meat (dark, firm, and dry) occurs if the glycogen reserves, due to challenges, are already used up, and the lactic acid production is insufficient. Especially PSE meat is closely related to breeds – some are more susceptible to stress, others less. However, some trials show that influencing pH in meat is possible to a certain extent.

He et al., 2022 added yeast ß-glucan to the diets of finishing pigs and a higher pH45 min (linear and quadratic, p<0.01) and a higher redness (a*; linear, p<0.05) of the meat. Wu et al. (2020) achieved a significantly increased pH24h through the addition of Glucose oxidase produced by Aspergillus niger.

Sensory characteristics are very subjective

In general, the sensory characteristics of meat are seen very individually. Some prefer lean, others fatty meat, some like meat with a characteristic taste, and others with a neutral. However, the typical meat taste of umami is partly determined by the nucleotide inosine monophosphate (IMP), which is regarded as an essential index for evaluating meat flavor and the acceptability of meat products. IMP provides about 40-fold higher umami taste than sodium glutamate (Huang et al. 2022).IMP is the organophosphate of inosin. Inosine, however, according to Kroemer and Zitvogel (2020), is produced by Bifidobacterium pseudolongum, which possibly can be controlled by feeding. Sun et al. (2018) compared Caoke and Partridge Shank chickens and divided them into free-range and cage groups. They found out that, except for acids, the amounts of flavor components were higher in the free-range than in the cage groups. The two housing systems also modified the microbiota, and Sun et al. took it as an indication that meat flavor, as well as the composition and diversity of gut microbiota, are closely associated with the housing systems. Fu et al. (2023) examined the addition of a mixture containing Pulsatilla, Gentian, and Rhizoma coptidis and a mixture with Codonopsis pilosula, Atractylodes, Poria cocos, and Licorice to the feed of Hungarian white geese. They saw that in both groups, the total amino acid levels, especially Glu, Lys, and Asp, increased, with, according to Liu et al. (2018), Glu and Asp directly affecting meat’s freshness and flavor. Yu et al. (2021) achieved similar results by adding resveratrol to the diet of Peking ducks. The addition of the herbs additionally led to a higher Firmicutes/Bacteroidetes ratio and an increased level of lactobacilli (Fu et al., 2023).

How can EW Nutrition’s feed additives help to improve meat quality?

Meat quality is influenced by the microbiome. So, feed additives that stabilize the microbiome or promote certain beneficial bacterial strains are an opportunity.

Ventar D modulates the microbiome

Ventar D balances the microbiome by promoting beneficial bacteria such as lactobacilli and fighting harmful ones such as Clostridia, E. coli, and Salmonella. (Heinzl, 2022). In another trial with broilers, the addition of Ventar D to all feeds (100 g/t) showed an increase in short-chain fatty acids in the intestine:

Figure Short Chain Fatty AcidsFigure 1: Short-chain fatty acids in the cecum of broilers

Santoquin countersteers oxidation

Another helpful product category is antioxidants. They can prevent the oxidation of lipids and proteins. For this purpose, EW Nutrition offers Santoquin M6*, a product tested by Kuttapan et al. (2021). Santoquin M6 was tested concerning its ability to minimize the oxidative damage caused by feeding oxidized fat. A control group receiving oxidized fat in feed was compared to one receiving oxidized fat plus 188 ppm Santoquin M6 (≙125 ppm ethoxyquin). The main parameters for this study were TBARS in the breast muscle, the incidence of wooden breast, and the live weight on day 48.

Results indicated that the inclusion of Santoquin M6 reduced the production of TBARS in the breast muscles, demonstrating a lower level of oxidative stress in the breast muscles.

Figure Breast Muscle TBARSFigure 2: Thiobarbituric acid reactive substances (TBARS) in broiler breast muscles. TBARS are formed as a by-product of lipid peroxidation.

Additionally, it reduced the incidence of severe woody breasts (Score 3) by almost half and helped mitigate the impact of breast muscle degradation due to increased oxidative stress.

Figure Incidence Of Wooden BreastFigure 3: Incidence of wooden breast in broilers

*Usage of ethoxyquin is dependent on country regulations.

Feed hygiene with Acidomix products minimizes harmful pathogens

The Acidomix product line offers liquid, powdery, and micro-granulated products to be added to feed and water. The organic acids in Acidomix directly act against pathogens in the feed and the water and help keep the intestinal flora in balance.

A trial evaluating the effect of different Acidomix products against diverse pathogens showed lower MICs for most Acidomix products than for single organic acids. The trial was conducted with decreasing concentrations of the Acidomix products (2 – 0.015625 %) and 105 CFU of the respective microorganisms (microtiter plates; 50 µl bacterial solution and 50 µl diluted product).

Figure Minimum Inhibiting Concentration
Feeding is the one side, slaughtering the other one

With feeding, the microbiota and some meat characteristics can be changed; however, the last step, handling the animals before and the meat after slaughtering also significantly contributes to a good quality of meat. Stress due to the transport and the slaughterhouse atmosphere, combined with stress-sensible breeds, can lead to PSE meat. Incorrect handling at the slaughterhouse can lead to meat contaminated with pathogens.

Combining feeding measures with professional and calm handling of the animals is the best strategy to achieve high-quality meat.

 

References

Aaslyng, Margit D., and Lene Meinert. “Meat Flavour in Pork and Beef – from Animal to Meal.” Meat Science 132 (2017): 112–17. https://doi.org/10.1016/j.meatsci.2017.04.012.

Alfaig, Ebrahim, Maria Angelovičova, Martin Kral, and Ondrej Bučko. “EFF ECT of Probiotics and Thyme Essential Oil on the Essential Amino Acid Content of the Broiler Chicken Meat.” Acta Scientiarum Polonorum Technologia Alimentaria 13, no. 4 (2014): 425–32. https://doi.org/10.17306/j.afs.2014.4.9.

Allen, CD, SM Russell, and DL Fletcher. “The Relationship of Broiler Breast Meat Color and Ph to Shelf-Life and Odor Development.” Poultry Science 76, no. 7 (1997): 1042–46. https://doi.org/10.1093/ps/76.7.1042.

Balasubramanian, Balamuralikrishnan, Sang In Lee, and In-Ho Kim. “Inclusion of Dietary Multi-Species Probiotic on Growth Performance, Nutrient Digestibility, Meat Quality Traits, Faecal Microbiota, and Diarrhoea Score in Growing–Finishing Pigs.” Italian Journal of Animal Science 17, no. 1 (2017): 100–106. https://doi.org/10.1080/1828051x.2017.1340097.

Berri, Céile. “Variability of Sensory and Processing Qualities of Poultry Meat.” World’s Poultry Science Journal 56, no. 3 (2000): 209–24. https://doi.org/10.1079/wps20000016.

Cabling, M. M., H. S. Kang, B. M. Lopez, M. Jang, H. S. Kim, K. C. Nam, J. G. Choi, and K. S. Seo. “Estimation of Genetic Associations between Production and Meat Quality Traits in Duroc Pigs.” Asian-Australasian Journal of Animal Sciences 28, no. 8 (2015): 1061–65. https://doi.org/10.5713/ajas.14.0783.

Chen, Binlong, Diyan Li, Dong Leng, Hua Kui, Xue Bai, and Tao Wang. “Gut Microbiota and Meat Quality.” Frontiers in Microbiology 13 (2022). https://doi.org/10.3389/fmicb.2022.951726.

Chen, J.L., G.P. Zhao, M.Q. Zheng, J. Wen, and N. Yang. “Estimation of Genetic Parameters for Contents of Intramuscular Fat and Inosine-5′-Monophosphate and Carcass Traits in Chinese Beijing-You Chickens.” Poultry Science 87, no. 6 (2008): 1098–1104. https://doi.org/10.3382/ps.2007-00504.

Ding, Rongrong, Ming Yang, Jianping Quan, Shaoyun Li, Zhanwei Zhuang, Shenping Zhou, Enqin Zheng, et al. “Single-Locus and Multi-Locus Genome-Wide Association Studies for Intramuscular Fat in Duroc Pigs.” Frontiers in Genetics 10 (2019). https://doi.org/10.3389/fgene.2019.00619.

Dinh, Thu T., K. Virellia To, and M. Wes Schilling. “Fatty Acid Composition of Meat Animals as Flavor Precursors.” Meat and Muscle Biology 5, no. 1 (2021). https://doi.org/10.22175/mmb.12251.

Džinić, N., N. Puvača, T. Tasić, P. Ikonić, and Okanović. “How Meat Quality and Sensory Perception Is Influenced by Feeding Poultry Plant Extracts.” World’s Poultry Science Journal 71, no. 4 (2015): 673–82. https://doi.org/10.1017/s0043933915002378.

Džinić, N., N. Puvača, T. Tasić, P. Ikonić, and Okanović. “How Meat Quality and Sensory Perception Is Influenced by Feeding Poultry Plant Extracts.” World’s Poultry Science Journal 71, no. 4 (2015): 673–82. https://doi.org/10.1017/s0043933915002378.

Fu, Guilin, Yuxuan Zhou, Yupu Song, Chang Liu, Manjie Hu, Qiuyu Xie, Jingbo Wang, et al. “The Effect of Combined Dietary Supplementation of Herbal Additives on Carcass Traits, Meat Quality, Immunity and Cecal Microbiota Composition in Hungarian White Geese (v0.2)”.” Peer J.; 11:e15316, May 8, 2023. https://doi.org/10.7287/peerj.15316v0.2/reviews/3.

Fu, Guilin, Yuxuan Zhou, Yupu Song, Chang Liu, Manjie Hu, Qiuyu Xie, Jingbo Wang, et al. “The Effect of Combined Dietary Supplementation of Herbal Additives on Carcass Traits, Meat Quality, Immunity and Cecal Microbiota Composition in Hungarian White Geese.” PeerJ 11 (2023). https://doi.org/10.7717/peerj.15316.

Goodrich, Julia K., Jillian L. Waters, Angela C. Poole, Jessica L. Sutter, Omry Koren, Ran Blekhman, Michelle Beaumont, et al. “Human Genetics Shape the Gut Microbiome.” Cell 159, no. 4 (2014): 789–99. https://doi.org/10.1016/j.cell.2014.09.053.

Hazrati, S., V. Rezaeipour, and S. Asadzadeh. “Effects of Phytogenic Feed Additives, Probiotic and Mannan-Oligosaccharides on Performance, Blood Metabolites, Meat Quality, Intestinal Morphology, and Microbial Population of Japanese Quail.” British Poultry Science 61, no. 2 (2019): 132–39. https://doi.org/10.1080/00071668.2019.1686122.

He, Linjuan, Jianxin Guo, Yubo Wang, Lu Wang, Doudou Xu, Enfa Yan, Xin Zhang, and Jingdong Yin. “Effects of Dietary Yeast β-Glucan Supplementation on Meat Quality, Antioxidant Capacity and Gut Microbiota of Finishing Pigs.” Antioxidants 11, no. 7 (2022): 1340. https://doi.org/10.3390/antiox11071340.

Heinzl, Inge. “Efficient Microbiome Modulation with Phytomolecules.” EW Nutrition, July 6, 2022. https://ew-nutrition.com/pushing-microbiome-in-right-direction-phytomolecules/.

Huang, Zengwen, Juan Zhang, Yaling Gu, Zhengyun Cai, Dawei Wei, Xiaofang Feng, and Chaoyun Yang. “Analysis of the Molecular Mechanism of Inosine Monophosphate Deposition in Jingyuan Chicken Muscles Using a Proteomic Approach.” Poultry Science 101, no. 4 (2022): 101741. https://doi.org/10.1016/j.psj.2022.101741.

Jiao, Anran, Hui Diao, Bing Yu, Jun He, Jie Yu, Ping Zheng, Yuheng Luo, et al. “Infusion of Short Chain Fatty Acids in the Ileum Improves the Carcass Traits, Meat Quality and Lipid Metabolism of Growing Pigs.” Animal Nutrition 7, no. 1 (2021): 94–100. https://doi.org/10.1016/j.aninu.2020.05.009.

Kallus, Samuel J., and Lawrence J. Brandt. “The Intestinal Microbiota and Obesity.” Journal of Clinical Gastroenterology 46, no. 1 (2012): 16–24. https://doi.org/10.1097/mcg.0b013e31823711fd.

Khan, Muhammad Issa, Cheorun Jo, and Muhammad Rizwan Tariq. “Meat Flavor Precursors and Factors Influencing Flavor Precursors—a Systematic Review.” Meat Science 110 (2015): 278–84. https://doi.org/10.1016/j.meatsci.2015.08.002.

Kroemer, Guido, and Laurence Zitvogel. “Inosine: Novel Microbiota-Derived Immunostimulatory Metabolite.” Cell Research 30, no. 11 (2020): 942–43. https://doi.org/10.1038/s41422-020-00417-1.

Kuttappan, Vivek A., Megharaja Manangi, Matthew Bekker, Juxing Chen, and Mercedes Vazquez-Anon. “Nutritional Intervention Strategies Using Dietary Antioxidants and Organic Trace Minerals to Reduce the Incidence of Wooden Breast and Other Carcass Quality Defects in Broiler Birds.” Frontiers in Physiology 12 (2021). https://doi.org/10.3389/fphys.2021.663409

Lei, Jiaqi, Yuanyang Dong, Qihang Hou, Yang He, Yujiao Lai, Chaoyong Liao, Yoichiro Kawamura, Junyou Li, and Bingkun Zhang. “Intestinal Microbiota Regulate Certain Meat Quality Parameters in Chicken.” Frontiers in Nutrition 9 (2022). https://doi.org/10.3389/fnut.2022.747705.

Liu, R., M. Zheng, J. Wang, H. Cui, Q. Li, J. Liu, G. Zhao, and J. Wen. “Effects of Genomic Selection for Intramuscular Fat Content in Breast Muscle in Chinese Local Chickens.” Animal Genetics 50, no. 1 (2018): 87–91. https://doi.org/10.1111/age.12744.

Lo, L. L., D. G. McLaren, F. K. McKeith, R. L. Fernando, and J. Novakofski. “Genetic Analyses of Growth, Real-Time Ultrasound, Carcass, and Pork Quality Traits in Duroc and Landrace Pigs: II. Heritabilities and Correlations.” Journal of Animal Science 70, no. 8 (1992): 2387–96. https://doi.org/10.2527/1992.7082387x.

Luna, A., M.C. Lábaque, J.A. Zygadlo, and R.H. Marin. “Effects of Thymol and Carvacrol Feed Supplementation on Lipid Oxidation in Broiler Meat.” Poultry Science 89, no. 2 (2010): 366–70. https://doi.org/10.3382/ps.2009-00130.

Ma, Cui, Md. Abul Azad, Wu Tang, Qian Zhu, Wei Wang, Qiankun Gao, and Xiangfeng Kong. “Maternal Probiotics Supplementation Improves Immune and Antioxidant Function in Suckling Piglets via Modifying Gut Microbiota.” Journal of Applied Microbiology 133, no. 2 (2022): 515–28. https://doi.org/10.1111/jam.15572.

Ma, Jianfeng, Jingyun Chen, Mailin Gan, Lei Chen, Ye Zhao, Yan Zhu, Lili Niu, Shunhua Zhang, Li Zhu, and Linyuan Shen. “Gut Microbiota Composition and Diversity in Different Commercial Swine Breeds in Early and Finishing Growth Stages.” Animals 12, no. 13 (2022): 1607. https://doi.org/10.3390/ani12131607.

MacLeod, G. “The Flavour of Beef.” Essay. In Shahidi, F. (Eds) Flavor of Meat and Meat Products, 4–37. Boston, MA: Springer, 1994.

Mottram, Donald. “Flavour Formation in Meat and Meat Products: A Review.” Food Chemistry 62, no. 4 (1998): 415–24. https://doi.org/10.1016/s0308-8146(98)00076-4.

Okruszek, A. “Fatty Acid Composition of Muscle and Adipose Tissue of Indigenous Polish Geese Breeds.” Archives Animal Breeding 55, no. 3 (2012): 294–302. https://doi.org/10.5194/aab-55-294-2012.

Pereira, Ana Lúcia F., and Virgínia Kelly G. Abreu. “Lipid Peroxidation in Meat and Meat Products.” Essay. In Lipid Peroxidation Research. London: IntechOpen, 2020.

Puvača, Nikola, Tatjana Peulić, Predrag Ikonić, Sanja Popović, Jasmina Lazarević, Olivera Đuragić, Magdalena Cara, and Nedeljka Nikolova. “Effects of Medicinal Plants in Broiler Chicken Nutrition on  Selected Parameters of Meat Quality.” Macedonian Journal of Animal Science 9, no. 2 (2019): 45–51. https://doi.org/10.54865/mjas1992045p.

Richards, J. D., J. Gong, and C. F. de Lange. “The Gastrointestinal Microbiota and Its Role in Monogastric Nutrition and Health with an Emphasis on Pigs: Current Understanding, Possible Modulations, and New Technologies for Ecological Studies.” Canadian Journal of Animal Science 85, no. 4 (2005): 421–35. https://doi.org/10.4141/a05-049.

Serra, Valentina, Giancarlo Salvatori, and Grazia Pastorelli. “Dietary Polyphenol Supplementation in Food Producing Animals: Effects on the Quality of Derived Products.” Animals 11, no. 2 (2021): 401. https://doi.org/10.3390/ani11020401.

Stewart, S.M., G.E. Gardner, P. McGilchrist, D.W. Pethick, R. Polkinghorne, J.M. Thompson, and G. Tarr. “Prediction of Consumer Palatability in Beef Using Visual Marbling Scores and Chemical Intramuscular Fat Percentage.” Meat Science 181 (2021): 108322. https://doi.org/10.1016/j.meatsci.2020.108322.

Sun, Jing, Yan Wang, Nianzhen Li, Hang Zhong, Hengyong Xu, Qing Zhu, and Yiping Liu. “Comparative Analysis of the Gut Microbial Composition and Meat Flavor of Two Chicken Breeds in Different Rearing Patterns.” BioMed Research International 2018 (2018): 1–13. https://doi.org/10.1155/2018/4343196.

Thorslund, Cecilie A.H., Peter Sandøe, Margit Dall Aaslyng, and Jesper Lassen. “A Good Taste in the Meat, a Good Taste in the Mouth – Animal Welfare as an Aspect of Pork Quality in Three European Countries.” Livestock Science 193 (2016): 58–65. https://doi.org/10.1016/j.livsci.2016.09.007.

Vasquez, Robie, Ju Kyoung Oh, Ji Hoon Song, and Dae-Kyung Kang. “Gut Microbiome-Produced Metabolites in Pigs: A Review on Their Biological Functions and the Influence of Probiotics.” Journal of Animal Science and Technology 64, no. 4 (2022): 671–95. https://doi.org/10.5187/jast.2022.e58.

Wang, Cheng, Siyu Wei, Bojing Liu, Fengqin Wang, Zeqing Lu, Mingliang Jin, and Yizhen Wang. “Maternal Consumption of a Fermented Diet Protects Offspring against Intestinal Inflammation by Regulating the Gut Microbiota.” Gut Microbes 14, no. 1 (2022). https://doi.org/10.1080/19490976.2022.2057779.

Wen, Chaoliang, Qinli Gou, Shuang Gu, Qiang Huang, Congjiao Sun, Jiangxia Zheng, and Ning Yang. “The Cecal Ecosystem Is a Great Contributor to Intramuscular Fat Deposition in Broilers.” Poultry Science 102, no. 4 (2023): 102568. https://doi.org/10.1016/j.psj.2023.102568.

Werkhoff, Peter, Juergen Bruening, Roland Emberger, Matthias Guentert, Manfred Koepsel, Walter Kuhn, and Horst Surburg. “Isolation and Characterization of Volatile Sulfur-Containing Meat Flavor Components in Model Systems.” Journal of Agricultural and Food Chemistry 38, no. 3 (1990): 777–91. https://doi.org/10.1021/jf00093a041.

Wu, Choufei, Wentao Lyu, Qihua Hong, Xiaojun Zhang, Hua Yang, and Yingping Xiao. “Gut Microbiota Influence Lipid Metabolism of Skeletal Muscle in Pigs.” Frontiers in Nutrition 8 (2021). https://doi.org/10.3389/fnut.2021.675445.

Xiao, Yingping, Kaifeng Li, Yun Xiang, Weidong Zhou, Guohong Gui, and Hua Yang. “The Fecal Microbiota Composition of Boar Duroc, Yorkshire, Landrace and Hampshire Pigs.” Asian-Australasian Journal of Animal Sciences 30, no. 10 (2017): 1456–63. https://doi.org/10.5713/ajas.16.0746.

Yu, Qifang, Chengkun Fang, Yujing Ma, Shaoping He, Kolapo Matthew Ajuwon, and Jianhua He. “Dietary Resveratrol Supplement Improves Carcass Traits and Meat Quality of Pekin Ducks.” Poultry Science 100, no. 3 (2021): 100802. https://doi.org/10.1016/j.psj.2020.10.056.

Zhao, Guangmin, Yun Xiang, Xiaoli Wang, Bing Dai, Xiaojun Zhang, Lingyan Ma, Hua Yang, and Wentao Lyu. “Exploring the Possible Link between the Gut Microbiome and Fat Deposition in Pigs.” Oxidative Medicine and Cellular Longevity 2022 (2022): 1–13. https://doi.org/10.1155/2022/1098892.

Zhou, Xueyan, Xiaosong Jiang, Chaowu Yang, Bingcun Ma, Changwei Lei, Changwen Xu, Anyun Zhang, et al. “Cecal Microbiota of Tibetan Chickens from Five Geographic Regions Were Determined by 16s Rrna Sequencing.” MicrobiologyOpen 5, no. 5 (2016): 753–62. https://doi.org/10.1002/mbo3.367.

Zou, Xiaozhuo, Rong Xiao, Huali Li, Ting Liu, Yong Liao, Yuanliang Wang, Shusong Wu, and Zongjun Li. “Effect of a Novel Strain of Lactobacillus Brevis M8 and Tea Polyphenol Diets on Performance, Meat Quality and Intestinal Microbiota in Broilers.” Italian Journal of Animal Science 17, no. 2 (2017): 396–407. https://doi.org/10.1080/1828051x.2017.1365260.




Salmonella in pigs: a threat for humans and a challenge for pig producers

Header SWINE Fotolia

By Dr. Inge Heinzl, Editor, EW Nutrition

Salmonellosis is third among foodborne diseases leading to death (Ferrari, 2019). More than 91,000 human cases of Salmonellosis are reported by the EU each year, generating overall costs of up to €3 billion a year (EFSA, 2023), 10-20% of which are attributed to pork consumption (Soumet, 2022). The annual costs arising from the resulting human health losses in 2010 were about €90 million (FCC Consortium, 2010). Take the example of Ireland, where a high prevalence of Salmonella in lymph nodes still shows a severe issue pre-slaughter and a big challenge for slaughterhouses to stick to the process hygiene requirements (Deane, 2022).

Several governments already have monitoring programs in place, and the farms are categorized according to the salmonella contamination of their pigs. In some countries, e.g., Denmark, an economic penalty of 2% of the carcass value must be paid if the farm has level 2 (intermediate seroprevalence) and 4-8% if the level is 3. Other countries, e.g., Germany, the UK, Ireland, or the Netherlands, use quality assurance schemes. The farmers can only sell their carcasses under this label if their farm has a certain level.

Let’s take a quick look at the genus of Salmonella

Salmonellas are rod-shaped gram-negative bacteria of the family of enterobacteria that use flagella for their movement. They were named after the American vet Daniel Elmer Salmon. The genus of Salmonella consists of two species (S. bongori and S. enterica with seven subspecies) with in total more than 2500 serovars (see Figure 1). The effects of the different serovars can range from asymptomatic carriage to severe invasive systemic disease (Gal-Mor, 2014). All Salmonella serovars generally can cause disease in humans; the rosa-marked ones already showed infections.

Figure Genus Salmonella For ART PIGFigure 1: the genus of Salmonella with Salmonella serovars relevant for pigs (according to Bonardi, 2017: Salmonella in the pork production chain and its impact on human health in the European Union)

Within the group of Salmonella, some serovars can only reside in one or few species, e.g., S. enterica spp. enterica Serovar Dublin (S. Dublin) in bovines (Waldron, 2018) or S. Cholerasuis in pigs (Chiu, 2004). An infection in humans with these pathogens is often invasive and life-threatening (WHO, 2018). On the contrary, serovars like S. Typhimurium and S. Enteritidis are not host-specific and can cause disease in various species.

The serotypes S. Typhi and S. Paratyphi A, B, or C are highly adapted to humans and only for them pathogenic; they are responsible for the occurrence of typhus.

Serovars occurring in pigs and relevant for humans are, for example, S. Typhimurium (Hendriksen, 2004), S. Serotype 4,[5],12:I (Hauser et al., 2010), S. Cholerasuis (Chiu, 2004), S. Derby (Gonzalez-Santamarina, 2021), S. Agona (Brenner Michael, 2006) and S. Rissen (Elbediwi, 2021).

Transmission of Salmonella mostly happens via contaminated food

The way of transmission to humans depends on the serovar:
Human-specific and, therefore, only in humans and higher primates residing serovars S. Typhi and Paratyphi A, B, or C (typhoidal) are excreted via feces or urine. Therefore, any food or water contaminated with the feces or urine of infected people can transmit this disease (Government of South Australia, 2023). Typhoid and paratyphoid Salmonellosis occur endemic in developing countries with the lack of clean water and, therefore, inadequate hygiene (Gal-Mor, 2014).

Serovars which can cause disease in humans and animals (non-typhoidal), can be transmitted by
– animal products such as milk, eggs, meat
– contact with infected persons/animals (pigs, cows, pets, reptiles…) or
– other feces- or urine-contaminated products such as sprouts, vegetables, fruits….

Farm animals take salmonellas from their fellows, contaminated feed or water, rodents, or pests.

Symptoms of Salmonellosis can be severe

In the case of typhoid or paratyphoid Salmonellosis, the onset of illness is gradual. People can suffer from sustained high fever, unwellness, severe headache, and decreased appetite, but also from an enlarged spleen irritating the abdomen and dry cough.

A study conducted in Thailand with children suffering from enteric fever caused by the typhoid serovars S. Typhi and Paratyphi showed a sudden onset of fever and gastrointestinal issues (diarrhea), rose spots, bronchitis, and pneumonia (Thisyakorn et al., 1987)

The non-typhoid Salmonellosis is typically characterized by an acute onset of fever, nausea, abdominal pain with diarrhea, and sometimes vomiting (WHO, 2018). However, 5% of the persons – children with underlying conditions, e.g., babies, or people who have AIDS, malignancies, inflammatory bowel disease, gastrointestinal illness caused by non-typhoid serovars, and hemolytic anemia, or receiving an immunosuppressive therapy can be susceptible to bacteremia. Additionally, serovars like S. Cholerasuis or S. Dublin are apt to develop bacteremia by entering the bloodstream with little or no involvement of the gut (Chiu, 1999). In these cases, consequences can be septic arthritis, pneumonia, peritonitis, cutaneous abscess, mycotic aneurysm, and sometimes death (Chen et al., 2007; Chiu, 2004, Wang et al., 1996).

In pigs, S. Cholerasuis causes high fever, purple discolorations of the skin, and thereinafter diarrhea. The mortality rate in pigs suffering from this type of Salmonellosis is high. Barrows orally challenged with S. Typhimurium showed elevated rectal temperature by 12h, remaining elevated until the end of the study. Feed intake decreased with a peak at 48h after the challenge and remained up to 120h after the challenge. Daily gain reduced during the following two weeks after infection. A higher plasma cortisol level and a lower IGF-I level could also be noticed. All these effects indicate significant changes in the endocrine stress and the somatotropic axis, also without significant alterations in the systemic pro-inflammatory mediators (Balaji et al., 2000)

To protect humans, Salmonella in pork must be restraint

There are three main steps to keep the contamination of pork as low as possible:

  1. Keeping Salmonella out of the pig farm
  2. Minimizing spreading if Salmonella is already on the farm
  3. Minimizing contamination in the slaughterhouse

1. How to keep Salmonella out of the pig farm?

To answer this question, we must look at how the pathogen can be transported to the farm. According to the Code of Practice for the Prevention and Control of Salmonella on Pig Farms (Ministry of Agriculture, Fisheries and Food and the Scottish Executive Rural Affairs Department), there are several possibilities to infiltrate the pathogen into the farm:

  • Diseased pigs or pigs which are ill but don’t show any symptoms
  • Feeding stuff or bedding contaminated with dung
  • Pets, rodents, wild birds, or animals
  • Farm personnel or visitors
  • Equipment or vehicles

Caution with purchased animals!

To minimize/prevent the entry of Salmonella into the livestock, bought-in animals must come from reputable breeding farms with a salmonella monitoring system in place. As possible carrier animals are more likely to excrete Salmonella when stressed; they should be kept in isolation after purchasing. Additionally, the animals must go through a disinfectant foot bath before entering the farm.

Keep rodents, wild animals, and vermin in check!

Generally, the production site must be kept clean and as unattractive as possible for all these animals. Rests of feed must be removed, and dead animals and afterbirths must be promptly and carefully disposed of. A well-planned baiting and trapping policy should be in place to effectively control rodents.

Only selected people should enter the hog houses

In any case, the number of persons entering the hog house must be kept as low as possible. Farmworkers should be trained in the principles of hygiene. They should wear adequate clothing (waterproof boots and protective overalls) that can be easily cleaned/laundered and disinfected. The clothes/shoes should always be used only at this site. Thorough hand washing and the disinfection of the boots when entering and leaving the pig unit are a must.

If visits are necessary, the visitors should take the same measures as the farm workers. And, of course, they should not have had contact with another pig farm during the last 48 hours.

Keep pens, farm equipment, and vehicles clean!

Farm equipment should not be shared with other farms. If this cannot be avoided, it must be cleaned and disinfected before re-entering the farm. Also, the vehicles for the transport of the animals must be cleaned and disinfected as soon as possible after usage, as contaminated transporters always pose the risk of infection.

Feed should be Salmonella-free!

To get high feed quality, the feed should be purchased from feed mills/sources with a well-functioning bacterial control to guarantee the absence of Salmonella. It is essential that birds, domestic and wild animals cannot enter the feed stores.
It is also advised to keep dry feed dry as possibly contaminating Salmonella can multiply in such humid conditions. Additionally, all feed bins and delivery pipes for dry and wet feed must be consciously cleaned, and the damp feed pipes also disinfected.
The change from pellets to mash could be helpful as the pellets facilitate Salmonella colonization by stimulating the secretion of mucins (Hedemann et al., 2005).

For sanitation of the feed, we offer organic acids (Acidomix product range) or mixtures of organic acids and formaldehyde in countries where formaldehyde products are allowed (Formycine) to decrease the pathogenic load of the feed materials. In vitro trials show the effectiveness of the products:

 

For the in vitro trial with Formycine, autoclaved feed samples were inoculated with Salmonella enteritidis serovar Typhimurium DSM 19587 strain to reach a Salmonella contamination of 106 CFU/g of feed. After incubating at room temperature for three hours, Formycine Liquido was added to the contaminated feed samples at 0, 500, 1000, and 2000 ppm. The control and inoculated feed samples were further incubated at room temperature, and Salmonella counts (CFU/g) were carried out at 24, 48, 72 hours and on day 15. The limit of Salmonella detection was set at 100 CFU/g (102). Results are shown in figure 2.

Figure FormycineFig. 2: Effect of treatment time and different inclusion levels of Formycine Liquido on the Salmonella count in feed

As important as uncontaminated feed is clean water for drinking. It can be achieved by taking the water from a main or a bacteriologically controlled water borehole. Regular cleaning/disinfection of the tanks, pipes, and drinkers is essential.

Bedding should be Salmonella-free

Straw material containing feces of other animals (rodents, pets) always carries the risk of Salmonella contamination. Also, wet or moldy bedding is not recommended because it is an additional challenge for the animal. To optimize the quality of bedding, the straw should be bought from reliable and as few as possible sources. The material must be stored dry and as far as practicable from the pig buildings (Ministry of Agriculture, Fisheries and Food & Scottish Executive Rural Affairs Department, 2000).

Vaccination is a beneficial measure

For the control of Salmonella in swine herds, vaccination is an effective tool. De Ridder et al. (2013) showed that an attenuated vaccine reduced the transmission of Salmonella Typhimurium in pigs. The vaccination with an attenuated S. Typhimurium strain, followed by a booster vaccination with inactivated S. Cholerasuis, showed better effects than an inactivated S. Cholerasuis vaccine alone (Alborali et al., 2017). Bearson et al. (2017) could delimitate transmission through less shedding and protect the animals against systemic disease.
To achieve the best effects, the producer must understand the diversity of Salmonella serovars to choose the most promising vaccination strategy (FSIS, 2023).

2. How to minimize the spreading of Salmonella on the farm?

If there are already cases of Salmonella on the farm, infected animals must be separated from the rest of the herd. Small batch sizes are beneficial, as well as not mixing different litters after weaning. If feasible, separate units for different production phases with an all-in/all-out system could break the reinfection cycle and help reduce Salmonella contamination on the farm. And also in this case, vaccination is helpful.

Salmonella doesn’t like acid conditions

An effective tool is acidifying the feed with organic acids, as Salmonella doesn’t like acid conditions. A trial was conducted with Acidomix AFG and Acidomix AFL to show their effects against Salmonella. For the test, 105 CFU/g of Salmonella enterica ser. Typhimurium was added to feed containing 1000 ppm, 2000 ppm, and 3000 ppm of Acidomix AFG or AFL. The stomach and intestine were simulated in vitro by adjusting the pH with HCl and NaHCO3 as follows:
Stomach              2.8
Intestine              6.8-7.0

After the respective incubation, the microorganisms were recovered from feed and plated on an appropriate medium for CFU counting. The results are shown in figures 3 and 4.

Combi
Combi

Figures 3 + 4: Effects of different concentrations of Acidomix AFG and Acidomix AFL against Salmonella enterica ser. Typhimurium in feed

Phytomolecules can support pigs against Salmonella

Plant compounds or phytomolecules can also be used against Salmonella in pigs. Some examples of phytomolecules to be used are Piperine, Allicin, Eugenol, and Carvacrol. Eugenol, e.g., increases the permeability of the Salmonella membrane, disrupts the cytoplasmic membrane, and inhibits the production of bacterial virulence factors (Keita et al., 2022; Mak et al., 2019). Thymol and Carvacrol interact with the cell membrane by H bonding, also resulting in a higher permeability.

An already published in vitro trial conducted with our product Ventar D also showed excellent effects against Salmonella while sparing the beneficial gut flora. A further trial once more demonstrated the susceptibility of Salmonella to Ventar D. It showed that Ventar D controls Salmonella by suppressing their motility and, at higher concentrations, inactivating the cells (see figures 5 + 6):

Figure Motility TestFigure 5: S. enterica motility test: on the left side – control; on the right side – motility medium containing.750 µg/mL of Ventar
Figure Disk DiffusionFig 6 . Disk diffusion assay employing S. enterica. upper left side – disk containing 10 µL of Ventar; upper right – 5 µL; lower left – control; lower right – 1µL.

In addition to the direct Salmonella-reducing effect, essential oils / secondary plant compounds / phytomolecules improve digestive enzyme activity and digestion, leading to increased nutrient absorption and better feed conversion (Windisch et al., 2008).

3. How can the farmer keep Salmonella contamination low in the slaughterhouse?

In general, the slaughterhouse personnel is responsible for adequate hygiene management to prevent contamination of carcasses and meat. However, also the farmer can make his contribution to maintain the risk of contamination in the slaughterhouse as low as possible. A study by Vieira-Pinto (2006) revealed that one Salmonella-positive pig can contaminate several other carcasses.

According to a trial conducted by Hurd et al. (2002), infection and, therefore, “contamination” of other pigs can rapidly occur, meaning that cross-contamination is a topic during transport to the slaughterhouse and in the lairages when the pigs come together with animals from other farms. The stress to which the pigs are exposed influences physiological and biochemical processes. The microbiome and animal’s immunity are affected, leading to higher excretion of Salmonella during transport and in the lairages. So, the animals should not be stressed during loading and unloading or transportation. The trailer poses a further risk of infection if it was not cleaned and disinfected before. So, reliable people who treat the animals well and keep their trailers clean should be chosen for transportation.

Pig producers are obliged to keep Salmonella in check – phytomolecules can help

At least in the EU, pig producers have the big duty to keep Salmonella low in their herds; otherwise, they will have financial losses. They are not only responsible for their farm, but also the slaughterhouses count on them. Besides the standard strict hygiene management and vaccination, farmers can use products provided by the industry to sanitize feed but also to support their animals directly with phytomolecules acting against pathogens and supporting gut health.

All these measures together should be a solution to the immense challenge of Salmonella, to protect people and prevent economic losses.

References:

Alborali, Giovanni Loris, Jessica Ruggeri, Michele Pesciaroli, Nicola Martinelli, Barbara Chirullo, Serena Ammendola, Andrea Battistoni, Maria Cristina Ossiprandi, Attilio Corradi, and Paolo Pasquali. “Prime-Boost Vaccination with Attenuated Salmonella Typhimurium Δznuabc and Inactivated Salmonella Choleraesuis Is Protective against Salmonella Choleraesuis Challenge Infection in Piglets.” BMC Veterinary Research 13, no. 1 (2017): 284. https://doi.org/10.1186/s12917-017-1202-5.

Balaji, R, K J Wright, C M Hill, S S Dritz, E L Knoppel, and J E Minton. “Acute Phase Responses of Pigs Challenged Orally with Salmonella Typhimurium.” Journal of Animal Science 78, no. 7 (2000): 1885. https://doi.org/10.2527/2000.7871885x.

Bearson, Bradley L, Shawn M. Bearson, Brian W Brunelle, Darrell O Bayles, In Soo Lee, and Jalusa D Kich. “Salmonella Diva Vaccine Reduces Disease, Colonization, and Shedding Due to Virulent S. Typhimurium Infection in Swine.” Journal of Medical Microbiology 66, no. 5 (2017): 651–61. https://doi.org/10.1099/jmm.0.000482.

Brenner Michael, G, M Cardoso, and S Schwarz. “Molecular Analysis of Salmonella Enterica Subsp. Enterica Serovar Agona Isolated from Slaughter Pigs.” Veterinary Microbiology 112, no. 1 (2006): 43–52. https://doi.org/10.1016/j.vetmic.2005.10.011.

Chen, P.-L., C.-M. Chang, C.-J. Wu, N.-Y. Ko, N.-Y. Lee, H.-C. Lee, H.-I. Shih, C.-C. Lee, R.-R. Wang, and W.-C. Ko. “Extraintestinal Focal Infections in Adults with Non-typhoid Salmonella Bacteraemia: Predisposing Factors and Clinical Outcome.” Journal of Internal Medicine 261, no. 1 (2007): 91–100. https://doi.org/10.1111/j.1365-2796.2006.01748.x.

Chiu, Cheng-Hsun, Lin-Hui Su, and Chishih Chu. “Salmonella EntericaSerotype Choleraesuis: Epidemiology, Pathogenesis, Clinical Disease, and Treatment.” Clinical Microbiology Reviews 17, no. 2 (2004): 311–22. https://doi.org/10.1128/cmr.17.2.311-322.2004.

De Ridder, L., D. Maes, J. Dewulf, F. Pasmans, F. Boyen, F. Haesebrouck, E. Méroc, P. Butaye, and Y. Van der Stede. “Evaluation of Three Intervention Strategies to Reduce the Transmission of Salmonella Typhimurium in Pigs.” The Veterinary Journal 197, no. 3 (2013): 613–18. https://doi.org/10.1016/j.tvjl.2013.03.026.

Deane, Annette, Declan Murphy, Finola C. Leonard, William Byrne, Tracey Clegg, Gillian Madigan, Margaret Griffin, John Egan, and Deirdre M. Prendergast. “Prevalence of Salmonella spp. in Slaughter Pigs and Carcasses in Irish Abattoirs and Their Antimicrobial Resistance.” Irish Veterinary Journal 75, no. 1 (2022). https://doi.org/10.1186/s13620-022-00211-y.

Edel, W., M. Schothorst, P. A. Guinée, and E. H. Kampelmacher. “Effect of Feeding Pellets on the Prevention and Sanitation of Salmonella Infections in Fattening Pigs1.” Zentralblatt für Veterinärmedizin Reihe B 17, no. 7 (2010): 730–38. https://doi.org/10.1111/j.1439-0450.1970.tb01571.x.

EFSA. “Salmonella.” European Food Safety Authority. Accessed August 7, 2023. https://www.efsa.europa.eu/en/topics/topic/salmonella.

Elbediwi, Mohammed, Daiwei Shi, Silpak Biswas, Xuebin Xu, and Min Yue. “Changing Patterns of Salmonella Enterica Serovar Rissen from Humans, Food Animals, and Animal-Derived Foods in China, 1995–2019.” Frontiers in Microbiology 12 (2021). https://doi.org/10.3389/fmicb.2021.702909.

Elnekave, Ehud, Samuel Hong, Alison E Mather, Dave Boxrud, Angela J Taylor, Victoria Lappi, Timothy J Johnson, et al. “Salmonella Enterica Serotype 4,[5],12:I:- In Swine in the United States Midwest: An Emerging Multidrug-Resistant Clade.” Clinical Infectious Diseases 66, no. 6 (2018): 877–85. https://doi.org/10.1093/cid/cix909.

FCC Consortium. “Final Report – Food Safety.” European Commission, 2010. https://food.ec.europa.eu/system/files/2016-10/biosafety_food-borne-disease_salmonella_fattening-pigs_slaughthouse-analysis-costs.pdf.

Ferrari, Rafaela G., Denes K. Rosario, Adelino Cunha-Neto, Sérgio B. Mano, Eduardo E. Figueiredo, and Carlos A. Conte-Junior. “Worldwide Epidemiology of Salmonella serovars in Animal-Based Foods: A Meta-Analysis.” Applied and Environmental Microbiology 85, no. 14 (2019). https://doi.org/10.1128/aem.00591-19.

“FSIS Guideline to Control Salmonella in Swine Slaughter and Pork Processing Establishments.” FSIS Guideline to Control Salmonella in Swine Slaughter and Pork Processing Establishments | Food Safety and Inspection Service. Accessed August 14, 2023. https://www.fsis.usda.gov/guidelines/2023-0003.

Gal-Mor, Ohad, Erin C. Boyle, and Guntram A. Grassl. “Same Species, Different Diseases: How and Why Typhoidal and Non-Typhoidal Salmonella Enterica Serovars Differ.” Frontiers in Microbiology 5 (2014). https://doi.org/10.3389/fmicb.2014.00391.

González-Santamarina, Belén, Silvia García-Soto, Helmut Hotzel, Diana Meemken, Reinhard Fries, and Herbert Tomaso. “Salmonella Derby: A Comparative Genomic Analysis of Strains from Germany.” Frontiers in Microbiology 12 (2021). https://doi.org/10.3389/fmicb.2021.591929.

Government of South Australia. Typhoid and paratyphoid – including symptoms, treatment, and prevention, April 3, 2022. https://www.sahealth.sa.gov.au/wps/wcm/connect/public+content/sa+health+internet/conditions/infectious+diseases/typhoid+and+paratyphoid/typhoid+and+paratyphoid+-+including+symptoms+treatment+and+prevention.

Hauser, Elisabeth, Erhard Tietze, Reiner Helmuth, Ernst Junker, Kathrin Blank, Rita Prager, Wolfgang Rabsch, Bernd Appel, Angelika Fruth, and Burkhard Malorny. “Pork Contaminated with            Salmonella            Enterica            Serovar 4,[5],12:I:−, an Emerging Health Risk for Humans.” Applied and Environmental Microbiology 76, no. 14 (2010): 4601–10. https://doi.org/10.1128/aem.02991-09.

Health and Wellbeing; address=11 Hindmarsh Square, Adelaide scheme=AGLSTERMS.AglsAgent; corporateName=Department for. “Sa Health.” Typhoid and paratyphoid – including symptoms, treatment, and prevention, April 3, 2022. https://www.sahealth.sa.gov.au/wps/wcm/connect/public+content/sa+health+internet/conditions/infectious+diseases/typhoid+and+paratyphoid/typhoid+and+paratyphoid+-+including+symptoms+treatment+and+prevention.

Hedemann, M. S., L. L. Mikkelsen, P. J. Naughton, and B. B. Jensen. “Effect of Feed Particle Size and Feed Processing on Morphological Characteristics in the Small and Large Intestine of Pigs and on Adhesion of Salmonella Enterica Serovar Typhimurium DT12 in the Ileum in Vitro1.” Journal of Animal Science 83, no. 7 (2005): 1554–62. https://doi.org/10.2527/2005.8371554x.

Hendriksen, Susan W.M., Karin Orsel, Jaap A. Wagenaar, Angelika Miko, and Engeline van Duijkeren. “Animal-to-Human Transmission ofSalmonellaTyphimurium DT104A Variant.” Emerging Infectious Diseases 10, no. 12 (2004): 2225–27. https://doi.org/10.3201/eid1012.040286.

Keita, Kadiatou, Charles Darkoh, and Florence Okafor. “Secondary Plant Metabolites as Potent Drug Candidates against Antimicrobial-Resistant Pathogens.” SN Applied Sciences 4, no. 8 (2022). https://doi.org/10.1007/s42452-022-05084-y.

Ministry of Agriculture, Fisheries and Food, and Scottish Executive Rural Affairs Department. “Salmonella on Pig Farms – Code of Practice for the Prevention and Control Of.” ReadkonG.com, 2000. https://www.readkong.com/page/code-of-practice-for-the-prevention-and-control-of-5160969.

Morrow, W.E. Morgan, and Julie Funk. Ms. Salmonella as a Foodborne Pathogen in Pork. North Carolina State University Animal Science, n.d.

Soumet, C., A. Kerouanton, A. Bridier, N. Rose, M. Denis, I. Attig, N. Haddache, and C. Fablet. Report, Salmonella excretion level in pig farms and impact of quaternary ammonium compounds based disinfectants on Escherichia coli antibiotic resistance § (2022).

Thisyakorn, Usa. “Typhoid and Paratyphoid Fever in 192 Hospitalized Children in Thailand.” Archives of Pediatrics &amp; Adolescent Medicine 141, no. 8 (1987): 862. https://doi.org/10.1001/archpedi.1987.04460080048025.

Ung, Aymeric, Amrish Y. Baidjoe, Dieter Van Cauteren, Nizar Fawal, Laetitia Fabre, Caroline Guerrisi, Kostas Danis, et al. “Disentangling a Complex Nationwide Salmonella Dublin Outbreak Associated with Raw-Milk Cheese Consumption, France, 2015 to 2016.” Eurosurveillance 24, no. 3 (2019). https://doi.org/10.2807/1560-7917.es.2019.24.3.1700703.

Vieira-Pinto, M, R Tenreiro, and C Martins. “Unveiling Contamination Sources and Dissemination Routes of Salmonella Sp. in Pigs at a Portuguese Slaughterhouse through Macrorestriction Profiling by Pulsed-Field Gel Electrophoresis.” International Journal of Food Microbiology 110, no. 1 (2006): 77–84. https://doi.org/10.1016/j.ijfoodmicro.2006.01.046.

Waldron, P. “Keeping Cows and Humans Safe from Salmonella Dublin.” Cornell University College of Veterinary Medicine, December 25, 2018. https://www.vet.cornell.edu/news/20181218/keeping-cows-and-humans-safe-salmonella-dublin.

Wang, J.-H., Y.-C. Liu, M.-Y. Yen, J.-H. Wang, Y.-S. Chen, S.-R. Wann, and D.-L. Cheng. “Mycotic Aneurysm Due to Non-Typhi Salmonella: Report of 16 Cases.” Clinical Infectious Diseases 23, no. 4 (1996): 743–47. https://doi.org/10.1093/clinids/23.4.743.

WHO. “Salmonella (Non-Typhoidal).” World Health Organization, February 20, 2018. https://www.who.int/news-room/fact-sheets/detail/salmonella-(non-typhoidal).

Windisch, W., K. Schedle, C. Plitzner, and A. Kroismayr. “Use of Phytogenic Products as Feed Additives for Swine and Poultry1.” Journal of Animal Science 86, no. suppl_14 (2008). https://doi.org/10.2527/jas.2007-0459.

Windisch, W., K. Schedle, C. Plitzner, and A. Kroismayr. “Use of Phytogenic Products as Feed Additives for Swine and Poultry1.” Journal of Animal Science 86, no. suppl_14 (2008). https://doi.org/10.2527/jas.2007-0459.




Minimizing Collateral Effects of Antibiotic Administration in Swine Farms: A Balancing Act

PIGLETS FARM

By Dr Merideth Parke BVSc, Regional Technical Manager Swine, EW Nutrition

We care for our animals, and antibiotics are a crucial component in the management of disease due to susceptible pathogens, supporting animal health and welfare.  However, the administration of antibiotics in pig farming has become a common practice to prevent bacterial infections, reduce economic losses, and increase productivity.

All antibiotic applications have collateral consequences of significance, bringing a deeper consideration to their non-essential application. This article aims to challenge the choice to administer antibiotics by exploring the broader impact that antibiotics have on animal and human health, economies, and the environment.

Antibiotics disrupt microbial communities

Antibiotics do not specifically target pathogenic bacteria. By impacting beneficial microorganisms, they disrupt the natural balance of microbial communities within animals. They reduce the microbiota diversity and abundance of all susceptible bacteria – beneficial and pathogenic ones… many of which play crucial roles in digestion, brain function, the immune system, and respiratory and overall health. Resulting microbiota imbalances may present themselves in animals showing health performance changes associated with non-target systems, including the nasal, respiratory, or gut microbiome10, 9, 16. The gut-respiratory microbiome axis is well-established in mammals. Gut microbiota health, diversity, and nutrient supply directly impact respiratory health and function15. In pigs specifically, the modulation of the gut microbiome is being considered as an additional tool in the control of respiratory diseases such as PRRS due to the link between the digestion of nutrients, systemic immunity, and response to pulmonary infections12.

The collateral effect of antibiotic administration disrupting not only the microbial communities throughout the animal but also linked body systems needs to be considered significant in the context of optimal animal health, welfare, and productivity.

Antibiotic use can lead to the release of toxins

The consideration of the pathogenesis of individual bacteria is critical to mitigate potential for direct collateral effects associated with antibiotic administration. For example, in cases of toxin producing bacteria, when animals are medicated either orally or parenterally, mortality may increase due to the associated release of toxins when large numbers of toxin producing bacteria are killed quickly3.

Modulation of the brain function can be critical

Numerous animal studies have investigated the modulatory role of intestinal microbes on the gut-brain axis. One identified mechanism seen with antibiotic-induced changes in fecal microbiota is the decreased concentrations of hypothalamic neurotransmitter precursors, 5-hydroxytryptamine (serotonin), and dopamine6. Neurotransmitters are essential for communication between the nerve cells. Animals with oral antibiotic-induced microbiota depletion have been shown to experience changes in brain function, such as spatial memory deficits and depressive-like behaviors.

Processing of waste materials can be impacted

Anaerobic treatment technology is well accepted as a feasible management process for swine farm wastewater due to its relatively low cost with the benefit of bioenergy production. Additionally, the much smaller volume of sludge remaining after anaerobic processing further eases the safe disposal and decreases the risk associated with the disposal of swine waste containing residual antibiotics5.

The excretion of antibiotics in animal waste, and the resulting presence of antibiotics in wastewater, can impact the success of anaerobic treatment technologies, which already could be demonstrated by several studies8, 13. The degree to which antibiotics affect this process will vary by type, combination, and concentration. Furthermore, the presence of antibiotics within the anaerobic system may result in a population shift towards less sensitive microbes or the development of strains with antibiotic-resistant genes1, 14.

Antibiotics can be transferred to the human food chain

Regulatory authorities specify detailed withdrawal periods after antibiotic treatment. However, residues of antibiotics and their metabolites may persist in animal tissues, such as meat and milk, even after this period. These residues can enter the human food chain if not adequately monitored and controlled.

Prolonged exposure to low levels of antibiotics through the consumption of animal products may contribute to the emergence of antibiotic-resistant bacteria in humans, posing a significant public health risk.

Contamination of the environment

As already mentioned before, the administration of antibiotics to livestock can result in the release of these compounds into the environment. Antibiotics can enter the soil, waterways, and surrounding ecosystems through excretions from treated animals, inappropriate disposal of manure, and runoff from agricultural fields. Once in the environment, antibiotics can contribute to the selection and spread of antibiotic-resistant bacteria in natural bacterial communities. This contamination poses a potential risk to wildlife, including birds, fish, and other aquatic organisms, as well as the broader ecological balance of affected ecosystems.

Every use of antibiotics can create resistance

One of the widely researched concerns associated with antibiotic use in livestock is the development of antibiotic resistance. The development of AMR does not require prolonged antibiotic use and, along with other collateral effects, also occurs when antibiotics are used within recommended therapeutic or preventive applications.

Gene mutations can supply bacteria with abilities that make them resistant to certain antibiotics (e.g., a mechanism to destroy or discharge the antibiotic). This resistance can be transferred to other microorganisms, as seen with the effect of carbadox on Escherichia coli7 and Salmonella enterica2 and the carbadox and metronidazole effect on Brachyspira hyodysenteriae16. Additionally, there is an indication that the zinc resistance of Staphylococcus of animal origin is associated with the methicillin resistance coming from humans4.

Consequently, the effectiveness of antibiotics in treating infections in target animals becomes compromised, and the risk of exposure to resistant pathogens for in-contact animals and across species increases, including humans.

Alternative solutions are available

To successfully minimize the collateral effects of antibiotic administration in livestock, a unified strategy with support from all stakeholders in the production system is essential. The European Innovation Partnership – Agriculture11 concisely summarizes such a process as requiring…

  1. Changing human mindsets and habits: this is the first and defining step to successful antimicrobial reduction
  2. Improving pig health and welfare: Prevention of disease with optimal husbandry, hygiene, biosecurity, vaccination programs, and nutritional support.
  3. Effective antibiotic alternatives: for this purpose, phytomolecules, pro/pre-biotics, organic acids, and immunoglobulins are considerations.

In general, implementing responsible antibiotic stewardship practices is paramount. This includes limiting antibiotic use to the treatment of diagnosed infections with an effective antibiotic, and eliminating their use as growth promotors or for prophylactic purposes.

Keeping the balance is of crucial importance

While antibiotics play a crucial role in ensuring the health and welfare of livestock, their extensive administration in the agricultural industry has collateral effects that cannot be ignored. The development of antibiotic resistance, environmental contamination, disruption of microbial communities, and the potential transfer of antibiotic residues to food pose significant challenges.

Adopting responsible antibiotic stewardship practices, including veterinary oversight, disease prevention programs, optimal animal husbandry practices, and alternatives to antibiotics, can strike a balance between animal health, efficient productive performance, and environmental and human health concerns.

The collaboration of stakeholders, including farmers, veterinarians, policymakers, industry and consumers, is essential in implementing and supporting these measures to create a sustainable and resilient livestock industry.

References

  1. Angenent, Largus T., Margit Mau, Usha George, James A. Zahn, and Lutgarde Raskin. “Effect of the Presence of the Antimicrobial Tylosin in Swine Waste on Anaerobic Treatment.” Water Research 42, no. 10–11 (2008): 2377–84. https://doi.org/10.1016/j.watres.2008.01.005.
  2. Bearson, Bradley L., Heather K. Allen, Brian W. Brunelle, In Soo Lee, Sherwood R. Casjens, and Thaddeus B. Stanton. “The Agricultural Antibiotic Carbadox Induces Phage-Mediated Gene Transfer in Salmonella.” Frontiers in Microbiology 5 (2014). https://doi.org/10.3389/fmicb.2014.00052.
  3. Castillofollow, Manuel Toledo, Rocío García Espejofollow, Alejandro Martínez Molinafollow, María Elena  Goyena Salgadofollow, José Manuel Pintofollow, Ángela Gallardo Marínfollow, M. Toledo, et al. “Clinical Case: Edema Disease – the More I Medicate, the More Pigs Die!” $this->url_servidor, October 15, 2021. https://www.pig333.com/articles/edema-disease-the-more-i-medicate-the-more-pigs-die_17660/.
  4. Cavaco, Lina M., Henrik Hasman, Frank M. Aarestrup, Members of MRSA-CG:, Jaap A. Wagenaar, Haitske Graveland, Kees Veldman, et al. “Zinc Resistance of Staphylococcus Aureus of Animal Origin Is Strongly Associated with Methicillin Resistance.” Veterinary Microbiology 150, no. 3–4 (2011): 344–48. https://doi.org/10.1016/j.vetmic.2011.02.014.
  5. Cheng, D.L., H.H. Ngo, W.S. Guo, S.W. Chang, D.D. Nguyen, S. Mathava Kumar, B. Du, Q. Wei, and D. Wei. “Problematic Effects of Antibiotics on Anaerobic Treatment of Swine Wastewater.” Bioresource Technology 263 (2018): 642–53. https://doi.org/10.1016/j.biortech.2018.05.010.
  6. Köhler, Bernd, Helge Karch, and Herbert Schmidt. “Antibacterials That Are Used as Growth Promoters in Animal Husbandry Can Affect the Release of Shiga-Toxin-2-Converting Bacteriophages and Shiga Toxin 2 from Escherichia Coli Strains.” Microbiology 146, no. 5 (2000): 1085–90. https://doi.org/10.1099/00221287-146-5-1085.
  7. Loftin, Keith A., Cynthia Henny, Craig D. Adams, Rao Surampali, and Melanie R. Mormile. “Inhibition of Microbial Metabolism in Anaerobic Lagoons by Selected Sulfonamides, Tetracyclines, Lincomycin, and Tylosin Tartrate.” Environmental Toxicology and Chemistry 24, no. 4 (2005): 782–88. https://doi.org/10.1897/04-093r.1.
  8. Looft, Torey, Heather K Allen, Brandi L Cantarel, Uri Y Levine, Darrell O Bayles, David P Alt, Bernard Henrissat, and Thaddeus B Stanton. “Bacteria, Phages and Pigs: The Effects of in-Feed Antibiotics on the Microbiome at Different Gut Locations.” The ISME Journal 8, no. 8 (2014a): 1566–76. https://doi.org/10.1038/ismej.2014.12.
  9. Looft, Torey, Heather K. Allen, Thomas A. Casey, David P. Alt, and Thaddeus B. Stanton. “Carbadox Has Both Temporary and Lasting Effects on the Swine Gut Microbiota.” Frontiers in Microbiology 5 (2014b). https://doi.org/10.3389/fmicb.2014.00276.
  10. Nasralla, Meisoon. “EIP-Agri Concept.” EIP-AGRI – European Commission, September 11, 2017. https://ec.europa.eu/eip/agriculture/en/eip-agri-concept.html.
  11. Niederwerder, Megan C. “Role of the Microbiome in Swine Respiratory Disease.” Veterinary Microbiology 209 (2017): 97–106. https://doi.org/10.1016/j.vetmic.2017.02.017.
  12. Poels, J., P. Van Assche, and W. Verstraete. “Effects of Disinfectants and Antibiotics on the Anaerobic Digestion of Piggery Waste.” Agricultural Wastes 9, no. 4 (1984): 239–47. https://doi.org/10.1016/0141-4607(84)90083-0.
  13. Shimada, Toshio, Julie L. Zilles, Eberhard Morgenroth, and Lutgarde Raskin. “Inhibitory Effects of the Macrolide Antimicrobial Tylosin on Anaerobic Treatment.” Biotechnology and Bioengineering 101, no. 1 (2008): 73–82. https://doi.org/10.1002/bit.21864.
  14. Sikder, Md. Al, Ridwan B. Rashid, Tufael Ahmed, Ismail Sebina, Daniel R. Howard, Md. Ashik Ullah, Muhammed Mahfuzur Rahman, et al. “Maternal Diet Modulates the Infant Microbiome and Intestinal Flt3l Necessary for Dendritic Cell Development and Immunity to Respiratory Infection.” Immunity 56, no. 5 (May 9, 2023): 1098–1114. https://doi.org/10.1016/j.immuni.2023.03.002.
  15. Slifierz, Mackenzie Jonathan. “The Effects of Zinc Therapy on the Co-Selection of Methicillin-Resistance in Livestock-Associated Staphylococcus Aureus and the Bacterial Ecology of the Porcine Microbiota,” 2016.
  16. Stanton, Thaddeus B., Samuel B. Humphrey, Vijay K. Sharma, and Richard L. Zuerner. “Collateral Effects of Antibiotics: Carbadox and Metronidazole Induce VSH-1 and Facilitate Gene Transfer among Brachyspira HyodysenteriaeApplied and Environmental Microbiology 74, no. 10 (2008): 2950–56. https://doi.org/10.1128/aem.00189-08.



Fighting antimicrobial resistance with immunoglobulins

OLYMPUS DIGITAL CAMERA

By Lea Poppe, Regional Technical Manager On-Farm Solutions Europe, and Dr. Inge Heinzl, Editor

One of the ten global public health threats is antimicrobial resistance (AMR). Jim O’Neill predicted 10 million people dying from AMR annually by 2050 (O’Neill, 2016). The following article will show the causes of antimicrobial resistance and how antibodies from the egg could help mitigate the problem of AMR.

Global problem of AMR results from the incorrect use of antimicrobials

Antimicrobial substances are used to prevent and cure diseases in humans, animals, and plants and include antibiotics, antivirals, antiparasitics, and antifungals. The use of these medicines does not always happen consciously, partially due to ignorance and partially for economic reasons.

There are various possibilities for the wrong therapy

  1. The use of antibiotics against diseases that household remedies could cure. A recently published German study (Merle et al., 2023) confirmed the linear relationship between treatment frequency and resistant scores in calves younger than eight months.
  2. The use of antibiotics against viral diseases: antibiotics only act against bacteria and not against viruses. Flu, e.g., is caused by a virus, but doctors often prescribe an antibiotic.
  3. Using broad-spectrum antibiotics instead of determining an antibiogram and applying a specific antibiotic.
  4. A too-long treatment with antimicrobials so that the microorganisms have the time to adapt. For a long time, the only mistake you could make was to stop the antibiotic therapy too early. Today, the motto is “as short as possible”.

Let’s take the example of neonatal calf diarrhea, one of the most common diseases with a high economic impact. Calf diarrhea can be caused by a wide range of bacteria, viruses, or parasites. This infectious form can be a complication of non-infectious diarrhea caused by dietary, psychological, and environmental stress (Uetake, 2012). The pathogens causing diarrhea in calves can vary with the region. In Switzerland and the UK, e.g., rotaviruses and cryptosporidia are the most common pathogens, whereas, in Germany, E. coli is also one of the leading causes. To minimize the occurrence of AMR, it is always crucial to know which pathogen is behind the disease.

Prophylactic use of antibiotics is still a problem

  1. The use of low doses of antibiotics to promote growth. This use has been banned in the EU now for 17 years now, but in other parts of the world, it is still common practice. Especially in countries with low hygienic standards, antibiotics show high efficacy.
  2. The preventive use of antibiotics to help, e.g., piglets overcome the critical step of weaning or to support purchased animals for the first time in their new environment. Antibiotics reduce pathogenic pressure, decrease the incidence of diarrhea, and ensure the maintenance of growth.
  3. Within the scope of prophylactic use of antimicrobials, also group treatment must be mentioned. In veal calves, group treatments are far more common than individual treatments (97.9% of all treatments), as reported in a study documenting medication in veal calf production in Belgium and the Netherlands. Treatment indications were respiratory diseases (53%), arrival prophylaxis (13%), and diarrhea (12%). On top, the study found that nearly half of the antimicrobial group treatment was underdosed (43.7%), and a large part (37.1%) was overdosed.

However, in several countries, consumers request reduced or even no usage of antibiotics (“No Antibiotics Ever” – NAE), and animal producers must react.

Today’s mobility enables the spreading of AMR worldwide

Bacteria, viruses, parasites, and fungi that no longer respond to antimicrobial therapy are classified as resistant. The drugs become ineffective and, therefore, the treatment of disease inefficient or even impossible. All the different usages mentioned before offer the possibility that resistant bacteria/microorganisms will occur and proliferate. Due to global trade and the mobility of people, drug-resistant pathogens are spreading rapidly throughout the world, and common diseases cannot be treated anymore with existing antimicrobial medicines like antibiotics. Standard surgeries can become a risk, and, in the worst case, humans die from diseases once considered treatable. If new antibiotics are developed, their long-term efficacy again depends on their correct and limited use.

Different approaches are taken to fight AMR

There have already been different approaches to fighting AMR. As examples, the annually published MARAN Report compiled in the Netherlands, the EU ban on antibiotic growth promoters in 2006, “No antibiotics ever (NAE) programs” in the US, or the annually published “Antimicrobial resistance surveillance in Europe” can be mentioned. One of the latest approaches is an advisory “One Health High-Level Expert Panel” (OHHLEP) founded by the Food and Agriculture Organization of the United Nations (FAO), the World Organization for Animal Health (OIE), the United Nations Environment Program (UNEP), and the World Health Organization (WHO) in May 2021. As AMR has many causes and, consequently, many players are involved in its reduction, the OHHLEP wants to improve communication and collaboration between all sectors and stakeholders. The goal is to design and implement programs, policies, legislations, and research to improve human, animal, and environmental health, which are closely linked. Approaches like those mentioned help reduce the spread of resistant pathogens and, with this, remain able to treat diseases in humans, animals, and plants.

On top of the pure health benefits, reducing AMR improves food security and safety and contributes to achieving the Sustainable Development Goals (e.g., zero hunger, good health and well-being, and clean water).

Prevention is better than treatment

Young animals like calves, lambs, and piglets do not receive immunological equipment in the womb and need a passive immune transfer by maternal colostrum. Accordingly, optimal colostrum management is the first way to protect newborn animals from infection, confirmed by 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 adequate supply.

Especially during the immunological gap when the maternal immunoglobulins are decreasing and the own immunocompetence is still not fully developed, it is crucial to have a look at housing, stress triggers, biosecurity, and the diet to reduce the risk of infectious diseases and the need for treatments.

Immunoglobulins from eggs additionally support young animals

Also, if newborn animals receive enough colostrum in time and if everything goes optimally, the animals suffer from two immunity gaps: the first one occurs just after birth before the first intake of colostrum, and the second one occurs when the maternal antibodies decrease, and the immune system of the young animal is still not developed completely. These immunity gaps raise the question of whether something else can be done to support newborns during their first days of life.

The answer was provided by Felix Klemperer (1893), a German internist researching immunity. He found that hens coming in contact with pathogens produce antibodies against these agents and transfer them to the egg. It is unimportant if the pathogens are relevant for chickens or other animals. In the egg, the immunoglobulins usually serve as an immune starter kit for the chick.

Technology enables us today to produce a high-value product based on egg powder containing natural egg immunoglobulins (IgY – immunoglobulins from the yolk). These egg antibodies mainly act in the gut. There, they recognize and tie up, for example, diarrhea-causing pathogens and, in this way, render them ineffective.

The efficacy of egg antibodies was demonstrated in different studies (Kellner et al., 1994; Erhard et al., 1996; Ikemori et al., 1997; Yokoyama et al., 1992; Marquart, 1999; Yokoyama et al., 1997) for piglets and calves.

Trial proves high efficacy of egg immunoglobulins in piglets

One trial conducted in Germany showed promising results concerning the reduction of mortality in the farrowing unit. For the trial, 96 sows and their litters were divided into three groups with 32 sows each. Two of the groups orally received a product containing egg immunoglobulins, the EP -1 + 3 group on days 1 and 3 and the EP – 1 + 2 + 3 group on the first three days. The third group served as a control. Regardless of the frequency of application, the egg powder product was very supportive and significantly reduced mortality compared to the control group. The measure resulted in 2 additionally weaned piglets than in the control group.

Figure

Egg immunoglobulins support young dairy calves

IgY-based products were also tested in calves to demonstrate their efficacy. In a field trial conducted on a Portuguese dairy farm with 12 calves per group, an IgY-containing oral application was compared to a control group without supplementation. The test product was applied on the day of birth and the two consecutive days. Key observation parameters during a two-week observation period were diarrhea incidence, onset, duration, and antibiotic treatments, the standard procedure on the trial farm in case of diarrhea. On-farm tests to check for the pathogenic cause of diarrhea were not part of the farm’s standards.

Figure

In this trial, 10 of 12 calves in the control group suffered from diarrhea, but in the trial group, only 5 calves. Total diarrhea and antibiotic treatment duration in the control group was 37 days (average 3.08 days/animal), and in the trial group, only 7 days (average 0.58 days/animal). Additionally, diarrhea in calves of the Globigen Calf Paste group started later, so the animals already had the chance to develop an at least minimally working immune system.

The supplement served as an effective tool to support calves during their first days of life and to reduce antibiotic treatments dramatically.

Conclusion

Antimicrobial reduction is one of the biggest tasks for global animal production. It must be done without impacting animal health and parameters like growth performance and general cost-efficacy. This overall demand can be supported with a holistic approach considering biosecurity, stress reduction, and nutritional support. Feed supplements such as egg immunoglobulins are commercial options showing great results and benefits in the field and making global animal production take the right direction in the future.

 

References upon request.




Respiratory challenges in pigs: Plants to the rescue!

Swine Pig Pixabay

By Dr. Inge Heinzl, Editor, EW Nutrition

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.



Global mycotoxin report: Jan-June 2022 | Find the pain points

myco map 22

By  Marisabel Caballero, Global Technical Manager Poultry, and Vinil Samraj Padmini, Global Category Manager Feed Quality, EW Nutrition 

The pressure of climate change is taking a severe toll – not just on weather-dependent industries, but already on society in general. For feed and food, the impact is already dramatic. Extreme weather events, increased temperatures, and rising carbon dioxide levels are facilitating the growth of toxigenic fungi in crops, severely increasing the risk of mycotoxin contamination. Once feed is contaminated, animal health can be impacted, with chain reactions affecting productivity for animal farming, as well as, ultimately, the quality and availability of food.

*** Download the full report for an analysis of mycotoxin contamination risks around the world




Acidifiers support piglets after weaning

8 piglet photo last page

By Dr. Inge Heinzl, Editor, EW Nutrition

In piglet production, high productivity, meaning high numbers of healthy and well-performing piglets weaned per sow and year, is the primary target. Diarrhea around weaning often gets in the way of achieving this goal.

Up to the ban of antibiotic growth promoters in 2006, antibiotics were often applied prophylactically to help piglets overcome this critical time. Zinc oxide (ZnO) application is another measure that cannot be used anymore to prevent piglet diarrhea. Effective alternatives are required.

Weaning – a critical point in piglets’ life

Weaning stress is well-known to have a negative impact on the balance of the intestinal microflora and gastrointestinal functions (Miller et al., 1985). Suckling piglets have a limited ability to produce hydrochloric acid, but nature has a solution to compensate for this inadequacy. The lactobacilli present in the stomach can use the lactose in the sow’s milk to produce lactic acid (Easter, 1988). In nature, the piglets would start to eat small amounts of solid feed at about three weeks when the sow’s milk production no longer covers their nutrient demand. By increasing the feed intake, the piglets stimulate hydrogen chloride (HCl) production in their stomachs.

In piglet production, where weaning occurs at three or four weeks of age, the piglets are still not eating considerable amounts of solid feed. It is often the case that 50 % of the piglets take feed at the earliest after 24 h, and 10 % accept the first feed only after 48 h (Brooks, 2001). Additionally, hard grains in the diet can physically damage the small intestine wall, reducing villus height and crypt depth (Kim et al., 2005).

Only a minor production of HCl, no more lactose supply for the lactobacilli, varying feed intake, and high buffering capacity of the feed lead to a pH of >5 in the stomach.

The higher stomach pH is partly responsible for problems after weaning

A pH higher than 5, besides causing direct effects on the microflora in the stomach, has consequences for the whole digestive tract and digestion.

A high pH is favorable for certain microorganisms, including coliforms (Sissons, 1989) and other acid-sensitive bacteria such as Salmonella typhimurium, Salmonella typhi, Campylobacter jejuni, and V. cholerae (Smith, 2003).

  1.  Lower activity of proteolytic enzymes

    In the stomach, the conversion of pepsinogen to pepsin, which is responsible for protein digestion, is catalyzed under acid conditions (Sanny et al., 1975). Pepsin works optimally at two pH levels: pH 2 and pH 3.5 (Taylor, 1959). With increasing pH, the activity decreases; at pH 6, it stops. Therefore, a high pH can lead to poor digestion and undigested protein arriving in the intestine. There, it can be used as “feed” for harmful bacteria, leading to their proliferation. Barrow et al. (1977) found higher counts of coliforms in piglets’ intestinal tract two days after weaning, while the number of lactobacilli was depressed.

    In the lower parts of the gastrointestinal tract (GIT), the final products of the pepsin protein digestion are needed to stimulate the secretion of pancreatic proteolytic enzymes. If no final products arrive, the enzymes are not activated, and the inadequate protein digestion continues. Additionally, gastric acid is the main stimulant for bicarbonate secretion in the pancreas, neutralizing gastric acid and providing an optimal pH environment for the digestive enzymes working in the duodenum.

  2. Expedited digesta transport

    The stomach pH also influences the transport of digesta. The acidity of the chyme leaving the stomach and arriving in the small intestine is decisive for the amount of digesta being transferred from the stomach to the small intestine. Acid-sensitive receptors provide feedback regulation to prevent the stomach from emptying until the duodenal chyme can be neutralized by pancreatic or other secretions (Pohl et al., 2008). Therefore, a higher pH in the stomach leads to a faster transport of the digesta, resulting in worse feed digestion.

  3. Proliferation of microorganisms

    A high pH is favorable for certain microorganisms, including coliforms (Sissons, 1989) and other acid-sensitive bacteria such as Salmonella typhimurium, Salmonella typhi, Campylobacter jejuni, and V. cholerae (Smith, 2003).

    Elevated stomach pH + incomplete immune system = diarrhea

Acidifiers can mitigate the adverse effects of weaning on piglets

To overcome this critical time of weaning and maintain performance, acidifiers can be a helpful tool. They improve gut health, stimulate immunity, and serve as nutrient sources – while also positively affecting feed and water hygiene.

What are acidifiers?

Acidifiers’ role in pig nutrition has evolved from feed preservatives to stomach pH stabilizers, compensating for young pigs’ reduced digestive capacity (Ferronato and Prandini, 2020). They are now used to replace antibiotic growth promoters and ZnO, which were applied for a long time to mitigate the negative effects of weaning.

In general, both organic and inorganic acids and their salts feature in animal nutrition. They can be added to the feed or the water.

Organic acids: Commonly used with good results

Feed acidifiers are usually organic acids, including fatty and amino acids. Their carboxyl functional group is responsible for their acidic specificity as feed additives (Pearlin et al.,2019). Their pKa, the pH where 50 % of the acid occurs in a dissociated form, is decisive for their antimicrobial action. In animal nutrition, acids with pKa 3-5 are typically used (Kirchgeßner and Roth, 1991).

Organic acids used as feed additives can be divided into three groups:

  •  Simple monocarboxylic acids such as formic, acetic, propionic, and butyric acid
  •  Carboxylic acids with a hydroxyl group such as lactic, malic, tartaric, and citric acid
  • Short-chain carboxylic acids with double bonds – fumaric and sorbic acid.

The primary acids for pig nutrition are acetic, fumaric, formic, lactic, benzoic, propionic, sorbic, and citric acids (Roth and Ettle, 2005).

Inorganic acids – the low-cost version

Inorganic acids are cheaper than organic acids, but their only effect is to decrease the pH. Additionally, they are extremely corrosive and dangerous liquids due to their strong acidity in a pure state (Kim et al., 2005).

Salts are easier to handle

The advantage of salts over free acids is that they are generally odorless and easier to handle in the feed manufacturing process due to their solid and less volatile form. Higher solubility in water is a further advantage compared to free acids (Huyghebaert and Van Immerseel, 2011; Roth and Ettle, 2005; Partanen and Mroz, 1999). The better handling and higher palatability make acid salts a more user-friendly method to apply acids to feed and water without compromising their efficacy (Luise et al., 2020).

The salts are mainly produced with calcium, potassium, and sodium. They include calcium formate, potassium diformate, sodium diformate, and sodium fumarate.

Blends

A mixture of diverse acidifiers combines the different characteristics of these substances. Perhaps, there may be synergistic effects. Acid blends are more and more used as feed additives. They have a wider-ranging action than single substances.

Roth et al. (1996) showed that a combination of formic acid with various formats is more effective than the application of formic acid alone.

The main effects of acidifiers

Acidifiers support piglets during the critical time after weaning through different modes of action. The final results are:

  • Improvement in gut health
  • Increase in growth performance
  • Stabilization of the immune system.

1.    Improvement in gut health

As shown in figure 1, the improvement in gut health relies on the antimicrobial effect of organic acids and the decrease in the stomach’s pH.

1.1     Organic acids directly attack bacteria

Organic acids not only act through their pH-decreasing effect but also directly attack pathogens. Due to their lipophilic character, organic acids can pass the bacterial cell membrane when they are in their undissociated form (Partanen in Piva et al., 2001). The lower the external pH, the more undissociated acid can pass the membrane.

Within the cell, the pH is higher. Hence, the organic acid dissociates and releases hydrogen ions, reducing the cytoplasmic pH from alkaline to acid. Cell metabolism is depressed at lower pH. Therefore, the bacterial cell needs to expel protons to get the cytoplasmic pH back to normal. As this is an energy-consuming process, more prolonged exposure to organic acids kills the bacterium. Additionally, the anions staying within the cell disturb the cell’s metabolic processes and participate in killing the bacterium.

Studies from Van Immerseel et al. (2006) revealed that many fermentative bacteria could let their intracellular pH decline and prevent increased acid penetration. Bacteria with a neutrophil pH, however, react more sensitively.

1.1     Decreased pH reduces non-acid-tolerant pathogens

There is a direct effect of pH on the microflora. Some pathogenic bacteria are susceptible to low pH. The proliferation of, e.g., E. coli, Salmonella, and Clostridium perfringens is minimized at a pH<5. Acid-tolerant bacteria such as lactobacilli or bifidobacteria, however, survive. Many lactobacilli can produce hydrogen peroxide, which inhibits, e.g., Staphylococcus aureus or Pseudomonas spp. (Juven and Pierson, 1996).

 

Already Fuller (1977) showed in in vitro experiments that certain bacteria such as Streptococci, Salmonella, and B. cereus don’t grow in an environment with pH 4.5 or even die (Micrococcus). In contrast, Lactobacilli are not so susceptible to this low pH. Using the same binding sites as harmful bacteria, they suppress coliforms, for example. Kirchgeßner et al. (1997) found a stronger reduction of E. coli than Lactobacilli and Bifidobacteria in different gut segments when exposed to 1.25 % formic acid.

1.2     Recovery of eubiosis through reduction of substrate

The reduction of the pH through organic acids maintains or stimulates the secretion of proteolytic enzymes in the stomach (pepsin) and pancreatic enzymes. Additionally, the acid leaving the stomach is partly responsible for regulating gastric emptying (Ravindran and Kornegay, 1993; Mayer, 1994). Both effects by improving protein digestion, reduce the fermentable substrates arriving in the hindgut. This decreases the quantity of fermentable substrate arriving in the intestine and, therefore, the growth of undesired pathogens.

2. Promotion of growth

2.1     Enhanced digestion of macronutrients

As explained above, the acidity in the stomach is responsible for the activation and stimulation of enzymes. Additionally, the lower pH keeps the feed in the stomach for longer. Both result in better digestion.

The improved utilization of nutrients leads to higher daily gain and better feed conversion. In pigs, the growth-promoting effect of organic acids is particularly pronounced during the first few weeks after weaning (Roth and Ettle, 2005). Some examples of the growth-promoting effect of formic and propionic acid feature in table 1.

Table 1: Influence of two commonly used organic acids in animals on growth performance

Varying results are mainly due to the character of the organic acid, the dosage, the buffering capacity, and the possible reduction of feed intake in case of a high dosage (Roth and Ettle, 2005).

2.2     Improved utilization of minerals

Minerals are essential for metabolic processes and, thus, healthy growth. Chelated minerals show a higher digestibility. Acidic anions of the acidifiers form complexes (chelates) with cationic minerals such as Ca, Zn, P, and Mg. The resulting higher digestibility and absorption lead to decreased excretion of supplemented minerals and, therefore, to a lower environmental burden. Kirchgeßner and Roth (1982), e.g., reported an improved absorption and retention of Ca, P, and Zn with the addition of fumaric acid. However, there are also trials showing no effect of acidification of the diet on mineral balance (Radecki et al., 1988).

Phytic acid

Another factor influencing the absorption of minerals, mainly phosphorus, is the amount of intrinsic or microbial phytase in the diet (Rutherfurd et al., 2012). The enzyme phytase releases phosphorus out of phytic acid and increases its bioavailability. Partanen and Mroz (1999) showed that organic acids improve the performance of phytase and, therefore, the bioavailability of phosphorus in the diet.

Besides a better utilization by the animal, improved absorption of minerals means preserving the environment and direct cost-saving, as mineral supplements are expensive.

2.3     Stimulation of gut and stomach mucosal morphology

An intact gut mucosa with a preferably high surface is vital for efficient nutrient absorption. Many trials show that organic acids improve the condition of the mucosa:

Organic acids stimulate cell proliferation

In an in vitro trial with pig hindgut mucosa, butyric acid stimulated epithelial cell proliferation in a dose-dependent manner (Sakata et al., 1995).

Blank et al. reported that fumaric acid, being a readily available energy source, may have a local trophic effect on the small intestines’ mucosa. Due to faster recovery of the gastrointestinal epithelial cells after weaning, this trophic effect may increase the absorptive surface and digestive capacity in the small intestines.

Organic acids influence villi length and crypt depth in the gut

Ferrara et al. (2016) observed a trend toward longer villi with a mixture of short-chain organic acids and mid-chain fatty acids, compared to the negative control.

The addition of Na-butyrate to the feed leads to increased crypt depth, villi length, and mucosa thickness in the distal jejunum and ileum, according to Kotunia et al. (2004). However, the villi length and mucosa thickness were reduced in the duodenum.

According to Gálfi and Bokori (1990), a diet with 0.17% sodium butyrate increased the length of ileal microvilli and the depth of caecal crypts in pigs weighing between 7 and 102 kg.

Organic acids strengthen stomach mucosa

Mazzoni et al. (2008) reported that sodium butyrate applied orally at a low dose influenced gastric morphology and function (thickening the mucosa), presumably due to its action on mucosal maturation and differentiation.

2.4    Pigs can use organic acids acid as an energy source

Organic acids are usually added to the feed in small doses. As some organic acids are intermediary products of the citric acid cycle, they are an energy source after being absorbed through the gut epithelium by passive diffusion. Their gross energy can be fully metabolized (Pearlin et al., 2019; Roth and Ettle, 2005; Suiryanrayna and Ramana, 2015).

The gross energy supply varies according to the acid. Roth and Ettle (2005) determined values between 6 kJ/g (formic acid) and 27 kJ/g (sorbic acid). Pearlin et al. (2019) calculated that 1 M of fumaric acid generates 1.340 kJ or 18 M ATP; this is comparable to the energy provision of glucose. Citric acid’s energy provision is similar; acetic and propionic acid require 18 and 15 % more energy to generate 1 M ATP.

Acidifiers improve immune response

The immune system, especially at the sensitive life stage of weaning, plays an essential role for the piglet. Acidifiers have been shown to stimulate or support the immune system. Ahmed et al. (2014) showed that citric acid (0.5 %) and a blend of acidifiers composed of formic, propionic, lactic, phosphoric acid + SO2(0.4 %) significantly increased the level of serum IgG. IgG are part of the humoral immune system. They mark foreign substances to be eliminated by other defense systems.


In a trial conducted by Ren et al. (2019), piglets receiving a mixture of formic and propionic acid showed lower concentrations of plasma tumor necrosis factor-α, regulating the activity of diverse immune cells. Furthermore, interferon-γ and interleukin (Il)-1ß were lower than in the control group after the challenge with E. coli (ETEC). In this trial, the addition of organic acids to the feed alleviated the inflammatory response in a way comparable to antibiotics.

In a nutshell

Organic acids are no longer seen as pure acidifiers but as growth promoters and potential antibiotic substitutes due to their positive effect on the gastrointestinal tract. Their main effect, the decrease of pH, entails consequences from inhibiting pathogenic bacteria and improved digestion to enhanced health and growth.
Research indicates that acidifiers can be a viable alternative to antibiotic growth promoters and ZnO for ensuring healthy piglet production after weaning.




Piglet performance with fewer antimicrobials is possible

veterinarianholdingdryfoodingranulesinhandsandoffering

By Technical Team, EW Nutrition

A variety of stressors simultaneously occur at weaning, making this probably the most challenging period in pig production. During weaning, we commonly see altered gut development and gut microbiome, which increases piglets’ vulnerability to diseases. The most classic clinical symptom resulting from these stressors is the occurrence of post-weaning diarrhea. It is a sign that something went wrong, and piglet development and overall performance may be compromised (Guevarra et al. 2019).

Besides weaning, an unavoidable practice in pig production, the swine industry has been facing other changes. Among them, the increased pressure to reduce the use of antimicrobials stands out. Antimicrobials are often associated with improved piglet performance and health. Their usage has been reduced worldwide, however, due to the threat of antimicrobial resistance that affects not just animal health but also human health (Cardinal et al., 2019).

Reduce antimicrobials and post-weaning diarrhea: can piglet nutrition achieve both?

With these drastic changes for the piglets and the global swine industry, producers must find solutions to keep their farms profitable — especially from a nutritional perspective. Our last article presented two feed additives that can be part of an antibiotic-free concept for post-weaning piglets. This article will highlight a few essential nutritional strategies that swine producers and nutritionists must consider when formulating post-weaning feed without or with reduced amounts of antimicrobials.

Pigs

What makes weaning so stressful for piglets?

Producers, nutritionists, and veterinarians all agree that weaning is a tough time for piglets (Yu et al., 2019) and, therefore, a challenge to all those involved in the pig production chain. Although there is a global trend towards increasing weaning age, generally speaking, animals are still immature when going through the weaning process. They face several physiological, nutritional, and environmental changes (figure 1).

Healthy Piglets
Figure 1. Factors associated with weaning can compromise piglet well-being and performance

Most of these changes become “stressors” that trigger a cascade of reactions affecting the balance and morphology of the intestinal microbiome (figure 2). The outcome is a decrease in the piglets’ well-being and, in most cases, performance. We need to clearly understand how these stressors affect pigs to develop effective strategies against post-weaning growth impairments, especially when no antimicrobials are allowed.

 

Schematic diagram
Figure 2. Schematic diagram illustrating the effects of stress in weaned piglets (adapted from Jayaraman and Nyachoti, 2017)

Weaning support starts before weaning

The use of creep feed has been evaluated and even criticized for many years. Some operations are still reluctant to use such a feed due to its high cost and amount of labor on the farm, with manually providing feed and cleaning feeding trays. In addition, some questions have been raised regarding the ideal composition of the creep feed – how much complexity should we add to this special diet?

Therefore, the benefits of creep feed are under re-evaluation, not only considering piglet physiology per se, but also feed characteristics and different feeding programs. Recent studies have questioned highly complex creep feed formulations. Creep feed is being called “transition feed” (Molist, 2021) – i.e., that meal which is complementary to sows’ milk and not a replicate of it, helping piglets during the period of changing its main source of nutrients. We must, therefore, look at it as a way of making piglets familiar with solid feed, as highlighted by Mike Tokach during the 2020 KSU Swine Day. Dr. Tokach also mentioned that the presence of feeders in the lactation pen could stimulate the exploratory behaviors of the piglets. Combined, these practices can lead to a higher feed intake and performance during the nursery phase.

Towards a pragmatic stance on creep feed

Heo et al. (2018) evaluated three different creep feed types: a highly digestible creep feed, weaning feed as creep feed, and sow feed as creep feed until weaning. Piglets receiving the highly digestible creep had higher feed intake during the second to the last week pre-weaning (14 to 21 days of age) and higher ADG during the last week pre-weaning (21 to 28 days of age). This resulted in a trend for higher weaning weight. However, these benefits did not persist after weaning when all piglets received the weaning feed.

Guevarra et al. (2019) also suggested that the abrupt transition in piglet nutrition to a more complex nutrient source can influence shifts in the gut microbiota, impacting the absorptive capacity of the small intestine. Yang et al. (2016) evaluated 40 piglets from eight litters during the first week after weaning. They found that the change in diet during weaning reduced the proliferation of intestinal epithelial cells. This indicates that this period affects cellular macromolecule organization and localization, in addition to energy and protein metabolism. These results suggest that “similarity” in feed pre- and post-weaning may contribute more to the continuity of nutrient intake post-weaning than a highly complex-nutrient dense creep feed.

Nutritional strategies without antibiotics: focus on pig physiology

As mentioned, it is crucial to avoid a drastic drop in feed/nutrient intake after weaning compared to pre-weaning levels. In a classic study, Pluske et al. (1996) showed the importance of high intake levels on villus weight (used as a reference for gut health, cf. graph 1). Although not desirable, the reduction should be considered “normal” behavior.

Imagine these recently weaned piglets, facing all these stressors, having to figure out within this new group of peers when it is time to eat, where to find food, why water and food now come from two distinct sources… Therefore, management, feeding, water quality, and other aspects play important roles in post-weaning feed intake (figure 3).

Average villus height
Graph 1. Villus height following different levels of feed intake (M = maintenance) post-weaning (a.b.c bars with unlike superscript letters are different at P<0.05). (From Pluske et al., 1996.)

From a nutritional perspective, piglets at weaning experience a transition from milk (a high-fat, low-carbohydrate liquid) to a plant-based diet (a solid, low-fat, and high-carbohydrate diet) (Guevarra et al., 2019). Even when previously introduced to solid feed, it is still difficult for their enzymatic system to cope with grains and beans.

One of the consequences of the lower digestibility capacity is an increase of undigested nutrients. Harmful bacteria thrive and cause diarrhea, reducing even further an already compromised feed intake. This cycle must be broken with the support of formulations based on piglet physiology.

Post-weaning feed must support digestion and nutrient absorption, including the largest possible share possible of high-quality, digestible ingredients, with low anti-nutritional factors. High-performing feed also integrates functional amino acids, functional carbohydrates, and additives to support the intestinal mucosa and gut microbiome.

Supporting piglets with effective solutions

Figure 3. Supporting piglets with effective solutions

Crude protein – more of the same?

Levels of crude protein in piglet feed have been in the spotlight for quite some time. The topic can be very controversial where the exact percentage of crude protein in the final feed is concerned. Some nutritionists pragmatically recommend maximal levels of 20% in the weaner feed. Others go a bit lower, with some formulations reaching 17 to 18% total crude protein. Levels above 20% will incur high costs and may accentuate the growth of pathogenic bacteria due to a higher amount of undigested protein in the distal part of the small intestine (figure 4).

crude protein levels in piglet feed
Figure 4. The dynamics of crude protein levels in piglet feed

What is not open for discussion, however, is the quality of the protein used, in terms of:

  • digestibility,
  • the total amount of anti-nutritional factors, and
  • the correct supply of essential and non-essential amino acids (particularly lysine, methionine, threonine, tryptophane, isoleucine and valine).

The critical role of digestibility

High-digestibility ingredients for piglets need to deliver minimum 85% digestibility. In most cases, to reach high biological values (correlating to high digestibility), these ingredients typically undergo different processing steps, including heat, physical, and chemical treatments. Animal by-products (such as hydrolyzed mucosa, fish meal, spray-dried plasma) and processed vegetable sources (soy protein concentrate, extruded grains, potato protein) can be used in high amounts during this phase. They will notably reduce the total amount of undigested protein reaching the distal part of the intestine, with 2 main benefits:

  • Less substrate for pathogenic bacteria proliferation (and therefore lower incidence of diarrhea)
  • Lower nitrogen excretion to the environment

 

Animal Feeds

It is common knowledge that certain storage proteins from soybean meal (for instance, glycinin and B-conglycinin) can cause damage to piglets’ intestinal morphology and trigger the activation of the immune system. However, it is normal practice to introduce this ingredient to piglets around weaning so that the animals can develop a certain level of tolerance to such compounds (Tokach et al., 2003). In Europe, where most diets are wheat-barley based, soybean meal is included in levels varying from 3 to 9% in the first 2 diets, with gradual increases during the nursery phase.

Amino acids and protein: manage the balance

When the supply and balance between essential and non-essential amino acids is concerned, reducing total crude protein brings indeed complexity to the formulations. The concept of ideal amino acid should be expanded, ideally, to all 9 essential amino acids (lysine, methionine, tryptophan, threonine, valine, isoleucine, leucine, histidine, and phenylalanine). In most cases, formulations go up to the 5th or 6th limiting amino acid. Lawor et al. (2020) suggest 2 practical approaches to avoid deficiencies when formulating low-protein piglet feed:

  • Maintain a maximum total lysine to crude protein ratio in the diet of 7.1 to 7.4%
  • Do not exceed the SID lysine to crude protein ratio of 6.4%

Some conditionally essential amino acids (e.g. arginine, proline, and glutamine) also play critical roles in diets with reduced crude protein levels. Glutamine is especially interesting. When supplemented in the feed, it can be used as a source of energy by the intestinal epithelium and, therefore, prevent atrophy and support nutrient absorption, resulting in better growth post-weaning (Hanczakowska and Niwińska, 2013; Watford et al., 2015)

The importance of the buffer capacity of the feed – supporting the enzymatic system

Given the move towards antibiotic reduction, this topic is more relevant than ever to nutritionists worldwide. The acid-binding capacity (also known as buffering capacity) of the feed directly affects the capacity of the stomach to digest protein. Hence, buffer capacity is of utmost importance in antimicrobial-free diets as it influences the growth of pathogenic bacteria (Lawlor et al., 2005).

In short, the acid-binding capacity is the resistance of an ingredient or complete feed to pH change. For piglet feed/feed ingredients, it is normally measured by the acid-binding capacity at pH4 (ABC-4). A higher ABC-4 equates to a higher buffering capacity. Feed with a high ABC-4 would require large amounts of gastric acid for the pH of the stomach to reach 4 and below. As the post-weaned piglet has limitations on producing and secreting acid, the stomach pH would stay high and, thus, less favorable for protein digestion.

The recommendation is to have a complete feed based on single ingredients with low ABC-4 values and to use additives that further reduce the ABC-4 value (such as organic acids). According to Molist (2020), post-weaning feed must have an ABC-4 that is lower than 250-300 meq/kg.

Talking about fiber

Dietary fibers are also known for regulating intestinal health in both humans and animals. Chen et al. (2020), for example, examined the effects of dietary soluble fibers (inulin) and insoluble fibers (lignocellulose) in weaned piglet diets for four weeks. Results showed that combining those fibers can positively influence nutrient digestibility, gut microbiota composition, intestinal barrier functions, and growth performance (table 1 ).

Effects of dietary fiber supplementation on piglet growth performance
Table 1. Effects of dietary fiber supplementation on piglet growth performance (adapted from Chen et al., 2020)

How to reduce antimicrobials? Understand the roles of piglet physiology and nutrition

Swine producers might think that “How can I reduce antimicrobial use on my farm?” and “How can I improve the performance of piglets at weaning?” are two separate questions. However, that is not always the case. Answers based on a deep understanding of physiology and nutrition dynamics help piglets overcome the challenges encountered during weaning – and, thus, lessen the need for antimicrobial interventions.

In this article, we have explored the basic principles that are the basis for ensuring the performance and health of the post-weaning piglet. Although we do not have a singular solution for eliminating antimicrobials on our pig farms, we can count on a group of robust and integrated nutritional strategies. By integrating factors ranging from management to feed additives, these solutions can improve piglet health and performance throughout their lives.

 

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INFOGRAPHIC: Healthy piglets after weaning

swine piglet kv

Piglet weaning is a critical period. When not properly managed, it leads to decreased performance, diarrhea, and sometimes mortality. 

The six areas of intervention in our infographic will help pig producers manage these stressors, avoid diarrhea, and maintain piglet health and performance. 

 

Piglets health and performance

 

 




The Zinc Oxide ban: What led to it, what are the alternatives?

shutterstock 1723596022 1 scaled

By Dr. Inge Heinzl, Editor, EW Nutrition

In June 2017, the European Commission decided to ban the use of veterinary drugs containing high doses of zinc oxide (3000mg/kg) from 2022. The use of zinc oxide in pig production must then be limited to a maximum level of 150ppm. Companies have been on the lookout for effective alternative strategies to maintain high profitability.

Modern pig production is characterised by its high intensity. In many European countries, piglets are weaned after 3-4 weeks, before their physiological systems are fully developed (e.g. immune and enzyme system). Weaning and thus separation from the mother, as well as a new environment with new germs, means stress for the piglets. Besides, the highly digestible sow’s milk, for which the piglets are wholly adapted, is replaced by solid starter feed.

This, associated with the above-mentioned stressors, can result in reduced feed intake during the first week after weaning and therefore in a delayed adaptation of the intestinal flora to the feed. Since the immune system of animals is not yet fully functional, pathogens such as enterotoxic E. coli can colonize the intestinal mucosa. This can possibly develop into a dangerous dysbiosis, leading to an increased incidence of diarrhea. Inadequate absorption results in suboptimal growth with worse feed conversion. The consequences are economic losses due to higher treatment costs, lower yields, and animal losses.

Diarrhea is one of the most common causes of economic losses in pig production. In the past, this was the reason antibiotics were prophylactically used as growth promoters. Antibiotics reduce antimicrobial pressure and have an anti-inflammatory effect. In addition to reducing the incidence of disease, they eliminate competitors for nutrients in the gut and thus improve feed conversion.

However, the use of antibiotics as growth promoters has been banned in the EU since 2006 due to increased antimicrobial resistance. As a result, zinc oxide (ZnO) appeared on the scene. A study carried out in Spain in 2012 (Moreno, 2012) showed that 57% of piglets received ZnO before weaning and 73% during the growth phase (27-75 days).

Zinc oxide: the disadvantages outweigh the advantages

What made the use of zinc oxide so attractive? Zinc oxide is inexpensive, available in many EU countries, and as a trace element it can be used in high doses through premixing. In some countries, however, a veterinary prescription is needed; in others, the use is already banned.

Zinc is a trace element involved in cell division and differentiation, and it influences the efficacy of enzymes. Since defence cells also need zinc, a supplementation that covers the demand for zinc strengthens the body’s defences. Through a positive effect on the structure of the gut mucosa membrane, zinc protects the body against the penetration of pathogenic germs.

If ZnO is used in pharmacological doses, it has a bactericidal effect against e.g. staphylococci (Ann et al., 2014) and various types of E. coli (Vahjen et al., 2016). Thus, prophylactic use prevents the incidence of diarrhea and the consequent decrease in performance. But the use of zinc oxide also has “side effects”.

Accumulation in the environment

Zinc belongs to the chemical group of heavy metals. For the use as a performance enhancer, it has to be administered in relatively high doses (2000–4000ppm). These high amounts are far above the physiological needs of the animals. With relatively low absorption rates (the bioavailability amounts to approximately 20% (European Commission, 2003)) and subsequent accumulation in manure, zinc can cause substantial contamination of the environment.

Encouraging the development of antibiotic resistance

In addition to the accumulation of zinc in the environment, another aspect also plays an important role: according to Vahjen et al. (2015), a dose of ≥2500mg/kg of food increases the presence of tetracycline and sulfonamide resistance genes in bacteria. In the case of Staphylococcus aureus, the development of resistance to zinc is combined with the development of resistance to methicillin (MRSA; Cavaco et al., 2011; Slifierz et al., 2015). A similar effect can be observed in the development of multiresistant E. coli (Bednorz et al., 2013; Ciesinski et al., 2018).  The reason for this is that the genes that encode antibiotic resistance, i.e. the ones that are “responsible” for the resistance, are found in the same plasmid (a DNA molecule that is small and independent of the bacterial chromosome).

Consequence: no more zinc oxide in the production of piglets from 2022 onwards

The negative effects on the environment and the promotion of antibiotic resistance led to the European Commission’s decision in 2017 to completely ban zinc oxide as a therapeutic agent and as a growth promoter in piglets within five years.

There are effective alternatives to zinc oxide

By the 2022 deadline, the EU pig industry must find a solution to replace ZnO. It must develop strategies that make future pig production efficient, even without substances such as antibiotics and zinc oxide. To this end, measures should be taken at different levels, such as farm management and biosecurity (e.g. effective hygiene management). The promotion of intestinal health for high animal performance is most important, however.

Promotion of gut health through stable gut microbiota

The term eubiosis denotes the balance of microorganisms living in a healthy intestine, which must be maintained to prevent diarrhea and ensure performance. However, weaning, food switching, and other external stressors can endanger this balance. As a result, potentially pathogenic germs can “overgrow” the commensal microbiome and develop dysbiosis. Through the use of functional supplements, intestinal health can be improved.

Phytomolecules – potent compounds created by nature

Phytomolecules, or secondary plant compounds, are substances formed by plants with a wide variety of properties. The best-known groups are probably essential oils, but there are also bitter substances, spicy substances, and other groups.

In animal nutrition, phytomolecules such as carvacrol, cinnamon aldehyde, and capsaicin can help improve intestinal health and digestion. They stabilize the intestinal flora by slowing or stopping the growth of pathogens that can cause disease. How? Phytomolecules, for example, make the cell walls of several bacteria permeable so that cell contents can leak. They also partially interfere with the enzymatic metabolism of the cell or intervene with the transport of ions, reducing the proton motive force. These effects depend on the dose: all these actions can destroy bacteria or at least prevent their proliferation.

What led to it, what are the alternatives?

Another point of attack for phytomolecules is the communication between microorganisms (quorum sensing). Phytomolecules can prevent microorganisms from releasing substances known as autoinducers, which they need to coordinate joint actions such as the formation of biofilms or the expression of virulence factors.

Medium-chain triglycerides and fatty acids

Medium-chain triglycerides (MCT) and fatty acids (MCFA) are characterised by a length of six to twelve carbon atoms. Thanks to their efficient absorption and metabolism, they can be optimally used as an energy source in piglet feeding. MCTs can be completely absorbed by the epithelial cells of the intestinal mucosa and hydrolysed with microsomal lipases. Hence they serve as an immediately available energy source and can improve the epithelial structure of the intestinal mucosa (Hanczakowska, 2017).

In addition, these supplements have a positive influence on the composition of the intestinal flora. Their ability to penetrate bacteria through semi-permeable membranes and destroy bacterial structures inhibits the development of pathogens such as salmonella and coliforms (Boyen et al., 2008; Hanczakowska, 2017; Zentek et al., 2011). MCFAs and MCTs can also be used very effectively against gram-positive bacteria such as streptococci, staphylococci, and clostridia (Shilling et al., 2013; Zentek et al., 2011).

Prebiotics

Prebiotics are short-chain carbohydrates that are indigestible for the host animal. However, certain beneficial microorganisms such as lactobacilli and bifidobacteria can use these substances as substrates. By selectively stimulating the growth of these bacteria, eubiosis is promoted (Ehrlinger, 2007). In pigs, mannan-oligosaccharides (MOS), fructooligosaccharides (FOS), inulin and lignocellulose are mainly used.

Another element of prebiotics’ positive effect on intestinal health is their ability to agglutinate pathogens. Pathogenic bacteria and MOS can bind to each other through lectin. This agglutination prevents pathogenic bacteria from adhering to the wall of the intestinal mucosa and thus from colonizing the intestine (Oyofo et al., 1989).

Probiotics

Probiotics can be used to regenerate an unbalanced gut flora. To do this, useful bacteria such as bifido or lactic acid bacteria are added to the food. They must settle in the gut and compete with the harmful bacteria.

There are also probiotics which target the communication between pathogens. In an experiment, Kim et al. (2017) found that the addition of probiotics that interfere with quorum sensing can significantly improve the microflora in weaned piglets and thus their intestinal health.

Organic acids

Organic acids show strong antibacterial activity in animals. In their undissociated form, the acids can penetrate bacteria. Inside, the acid molecule breaks down into a proton (H+) and an anion (HCOO-). The proton reduces the pH value in the bacterial cell and the anion interferes with the bacteria’s protein metabolism. As a result, bacterial growth and virulence are inhibited.

Conclusion

Today there are several possibilities in piglet nutrition to effectively support the young animals after weaning. The main objective is to maintain a balanced intestinal flora and therefore to sustain intestinal health – its deterioration often leads to diarrhea and hence to reduced returns. Intestinal health is promoted by stimulating beneficial bacteria and by inhibiting pathogenic ones. This can be achieved through feed additives that have an antibacterial effect and/or support the intestinal mucosa, such as phytomolecules, prebiotics, and medium-chain fatty acids. Through a combination of these possibilities, additive effects can be achieved. Piglets receive optimal support and the use of zinc oxide can be reduced.

 

References

Ann, Ling Chuo, Shahrom Mahmud, Siti Khadijah Mohd Bakhori, Amna Sirelkhatim, Dasmawati Mohamad, Habsah Hasan, Azman Seeni, and Rosliza Abdul Rahman. “Antibacterial Responses of Zinc Oxide Structures against Staphylococcus Aureus, Pseudomonas Aeruginosa and Streptococcus Pyogenes.” Ceramics International 40, no. 2 (March 2014): 2993–3001. https://doi.org/10.1016/j.ceramint.2013.10.008.

Bednorz, Carmen, Kathrin Oelgeschläger, Bianca Kinnemann, Susanne Hartmann, Konrad Neumann, Robert Pieper, Astrid Bethe, et al. “The Broader Context of Antibiotic Resistance: Zinc Feed Supplementation of Piglets Increases the Proportion of Multi-Resistant Escherichia Coli in Vivo.” International Journal of Medical Microbiology 303, no. 6-7 (August 2013): 396–403. https://doi.org/10.1016/j.ijmm.2013.06.004.

Boyen, F., F. Haesebrouck, A. Vanparys, J. Volf, M. Mahu, F. Van Immerseel, I. Rychlik, J. Dewulf, R. Ducatelle, and F. Pasmans. “Coated Fatty Acids Alter Virulence Properties of Salmonella Typhimurium and Decrease Intestinal Colonization of Pigs.” Veterinary Microbiology 132, no. 3-4 (December 10, 2008): 319–27. https://doi.org/10.1016/j.vetmic.2008.05.008.

Cavaco, Lina M., Henrik Hasman, Frank M. Aarestrup, Members Of Mrsa-Cg: Jaap A. Wagenaar, Haitske Graveland, Kees Veldman, et al. “Zinc Resistance of Staphylococcus Aureus of Animal Origin Is Strongly Associated with Methicillin Resistance.” Veterinary Microbiology 150, no. 3-4 (June 2, 2011): 344–48. https://doi.org/10.1016/j.vetmic.2011.02.014.

Ciesinski, Lisa, Sebastian Guenther, Robert Pieper, Martin Kalisch, Carmen Bednorz, and Lothar H. Wieler. “High Dietary Zinc Feeding Promotes Persistence of Multi-Resistant E. Coli in the Swine Gut.” Plos One 13, no. 1 (January 26, 2018). https://doi.org/10.1371/journal.pone.0191660.

Crespo-Piazuelo, Daniel, Jordi Estellé, Manuel Revilla, Lourdes Criado-Mesas, Yuliaxis Ramayo-Caldas, Cristina Óvilo, Ana I. Fernández, Maria Ballester, and Josep M. Folch. “Characterization of Bacterial Microbiota Compositions along the Intestinal Tract in Pigs and Their Interactions and Functions.” Scientific Reports 8, no. 1 (August 24, 2018). https://doi.org/10.1038/s41598-018-30932-6.

Ehrlinger, Miriam. 2007. “Phytogene Zusatzstoffe in der Tierernährung.“ PhD Diss., LMU München. URN: urn:nbn:de:bvb:19-68242.

European Commission. 2003. “Opinion of the Scientific Committee for Animal Nutrition on the use of zinc in feedingstuffs.”  https://ec.europa.eu/food/sites/food/files/safety/docs/animal-feed_additives_rules_scan-old_report_out120.pdf

Hanczakowska, Ewa. ”The use of medium chain fatty acids in piglet feeding – a review.” Annals of Animal Science 17, no. 4 (October 27, 2017): 967-977. https://doi.org/10.1515/aoas-2016-0099.

Hansche, Bianca Franziska. 2014. „Untersuchung der Effekte von Enterococcus faecium (probiotischer Stamm NCIMB 10415) und Zink auf die angeborene Immunantwort im Schwein. Dr. rer. Nat. Diss., Freie Universität Berlin. https://doi.org/10.17169/refubium-8548

Kim, Jonggun, Jaepil Kim, Younghoon Kim, Sangnam Oh, Minho Song, Jee Hwan Choe, Kwang-Youn Whang, Kwang Hyun Kim, and Sejong Oh. “Influences of Quorum-Quenching Probiotic Bacteria on the Gut Microbial Community and Immune Function in Weaning Pigs.” Animal Science Journal 89, no. 2 (November 20, 2017): 412–22. https://doi.org/10.1111/asj.12954.

Oyofo, Buhari A., John R. Deloach, Donald E. Corrier, James O. Norman, Richard L. Ziprin, and Hilton H. Mollenhauer. “Effect of Carbohydrates on Salmonella Typhimurium Colonization in Broiler Chickens.” Avian Diseases 33, no. 3 (1989): 531–34. https://doi.org/10.2307/1591117.

Shilling, Michael, Laurie Matt, Evelyn Rubin, Mark Paul Visitacion, Nairmeen A. Haller, Scott F. Grey, and Christopher J. Woolverton. “Antimicrobial Effects of Virgin Coconut Oil and Its Medium-Chain Fatty Acids On Clostridium Difficile.” Journal of Medicinal Food 16, no. 12 (December 2013): 1079–85. https://doi.org/10.1089/jmf.2012.0303.

Slifierz, M. J., R. Friendship, and J. S. Weese. “Zinc Oxide Therapy Increases Prevalence and Persistence of Methicillin-Resistant Staphylococcus Aureus in Pigs: A Randomized Controlled Trial.” Zoonoses and Public Health 62, no. 4 (September 11, 2014): 301–8. https://doi.org/10.1111/zph.12150.

Vahjen, Wilfried, Dominika Pietruszyńska, Ingo C. Starke, and Jürgen Zentek. “High dietary zinc supplementation increases the occurrence of tetracycline and sulfonamide resistance genes in the intestine of weaned pigs.” Gut Pathogens 7, article number 23 (August 26, 2015). https://doi.org/10.1186/s13099-015-0071-3.

Vahjen, Wilfried, Agathe Roméo, and Jürgen Zentek. “Impact of zinc oxide on the immediate postweaning colonization of enterobacteria in pigs.” Journal of Animal Science 94, supplement 3 (September 1, 2016): 359-363. https://doi.org/10.2527/jas.2015-9795.

Zentek, J., S. Buchheit-Renko, F. Ferrara, W. Vahjen, A.G. Van Kessel, and R. Pieper. “Nutritional and physiological role of medium-chain triglycerides and medium-chain fatty acids in piglets” Animal Health Research Reviews 12, no. 1 (June 2011): 83-93. https://doi.org/10.1017/s1466252311000089.