Enhancement of intestinal cell health and the resulting wellness of pets and livestock by the combined use of probiotics and butyric acid

Abstract

Probiotics, particularly Enterococcus faecium NCIMB 10415 strain named SF68 (registered trademark) as a probiotic active ingredient, are described to be used in combination with butyric acid or its salts to enhance the health of intestinal cells, and as a result, enhance the wellness of pets and livestock.

Description

Technical Field

[0001] The present invention relates to the field of animal nutrition, and more particularly to the use of probiotics in combination with butyric acid or its salts for promoting the health of intestinal cells and, as a result, the wellness of pets and livestock.

Background Art

[0002] Butyric acid and its salts, particularly sodium butyrate, have been used as animal feed for decades. For optimal use, animals need to absorb butyric acid or its salts through the uptake mechanism of this short-chain fatty acid (SCFA) into intestinal cells.

[0003] However, the uptake of butyric acid or its salts at the intestinal level may be impaired by inflammatory conditions caused by or characterized by damage to intestinal tissue and the epithelial barrier.

[0004] For example, in mice made obese by a high-fat diet, impaired uptake of butyric acid or its salts has been observed.

[0005] Therefore, it is necessary to optimally use butyric acid and its salts in animal feed, particularly for pets (dogs, cats) and livestock (fattening turkeys, chickens, laying hens, sows and piglets (nursing and weaning), fattening pigs, dairy cows, calves, etc.) to enhance the health of intestinal cells.

Summary of the Invention

[0006] Probiotics, particularly Enterococcus faecium strains, more particularly SF68® It has been found that by simultaneously administering to animals the Enterococcus faecium NCIMB 10415 strain named as a probiotic active ingredient, the uptake of butyric acid or its salts at the intestinal level of livestock and pets is improved.

[0007] Accordingly, an object of the present invention is to use Enterococcus faecium in combination with butyric acid or its salt as a feed supplement to enhance the health of intestinal cells, and as a result, enhance the wellness of pets and livestock.

BEST MODE FOR CARRYING OUT THE INVENTION

[0008] As used herein, "enhancing the health of intestinal cells" means enhancing the ability of intestinal cells to take up butyric acid. As a result, the wellness of the animal is improved, and from a practical point of view, it can mean an increase in the body weight of livestock and an improvement in physiological and behavioral parameters known as wellness indicators for pets, such as appetite, sleep, and the condition of the skin and fur. These wellness indicators in animals are well known to experts in this field.

[0009] Preferably, an Enterococcus faecium strain with accession number NCIMB 10415 named SF68 is used as the probiotic active ingredient. More preferably, SF68 is used in a microencapsulated form, such as those commercially available under the names Cervinet® LBC ME10, LBC ME20 plus, and LBC ME5 PET.

[0010] Butyric acid is preferably used in the form of a salt, particularly the sodium salt. Use in a microencapsulated form is particularly preferred, such as in a formulation commercially available under the name BUTiPEARL®.

[0011] In a preferred embodiment of the present invention, both the probiotic and butyric acid or its salt are mixed into a standard diet and given to the animal.

[0012] The amounts of the probiotic and butyric acid or its salt vary depending on the type of animal and the regulations of the region where it is applied.

[0013] For the sole purpose of providing information, and without limitation, examples of the dosages of butyrate (butyric acid) (ButiPEARL (registered trademark)) and SF68 (LBC ME10 and LBC ME20 plus for livestock, LBC ME5 PET for pets) are reported in Table 1 below.

Table 1

[0014] In its preferred embodiment, the present invention comprises feeding animals with a standard feed selected by experts in the field based on the type of animal and its breeding characteristics (such as dairy farming, meat consumption, etc.), to which the above-mentioned amount of butyric acid or its salt and the probiotic SF68, also selected based on the type of animal, are added.

[0015] Improved animal physiological growth and / or weight gain are obtained as compared to animals fed only with standard feed.

[0016] The idea of combining and using the probiotic SF68 with butyric acid or its salt is derived from experimental studies on mice treated with a high-fat diet, and it has been found that when administered together with SF68, butyric acid is better utilized / taken up by intestinal cells (see Example 1).

[0017] From the results of the mice, it was observed that SF68 normalizes the expression of the apical transporter of butyrate (butyric acid) altered by a high-fat diet (HFD).

[0018] In further studies on livestock, especially chickens, it was observed that the combined use of SF68 and butyric acid or its salt increased the body weight of the animals as compared to the control group fed only with standard feed.

[0019] This further feature of supplementing animal feed with butyric acid or its salt and the probiotic Enterococcus faecium (SF68) makes the present invention particularly advantageous.

[0020] Next, the usefulness, effectiveness, and advantages of the present invention will be described in more detail by the following examples, which do not limit the scope of the present invention in any way.

[0021] Example 1 SF60 reduces the butyrate concentration (butyric acid level) in the feces of high-fat diet (HFD)-fed mice Five-week-old C57BL / 6 mice (body weight 20 - 22 g) were provided by Envigo srl (Udine, Italy, San Pietro al Natisone). The mice were housed six per cage in a temperature-controlled room with a 12-hour light cycle at a temperature of 22 - 24°C and a humidity of 50 - 60% and acclimated for at least one week. The handling of the animals was in accordance with Directive 2010 / 63 / UE.

[0022] During the adaptation period, all mice were fed a standard diet (SD, 18% fat-derived calories; TD.2018). Thereafter, the animals were randomly divided into six groups of 10 mice each. - SD for 8 weeks - HFD for 8 weeks - SD + SF68(P) from week 4 (treatment 4 + 4) - HFD + SF68(P) from week 4 (treatment 4 + 4) - SD + SF68(P) for 8 weeks - HFD + SF68(P) for 8 weeks

[0023] The high-fat diet (HFD) had 60% calories from fat (TD.06414). A comparison of the calories of these two diets is reported below. JPEG2025519442000002.jpg20169

[0024] The body weight of the animals was measured once a week starting from the first day of the test. After 8 weeks, the animals were anesthetized and sacrificed. Blood samples and tissue samples were collected for further analysis and stored at -80°C.

[0025] For the analysis of short-chain fatty acids (SCFAs), fecal samples were freeze-dried and subjected to gas chromatography. The freeze-dried material was dissolved in 100 ml of 5 M formic acid and 400 ml of acetone and centrifuged (at 4000 rpm for 5 minutes). The SCFA concentration in the supernatant was measured using a GC2010 Plus gas chromatograph (Shimadzu Deutschland GmbH, Duisburg, Germany) equipped with a flame ionization detector and a thin-film capillary column Stabilwax® (Restek, Bad Homburg, Germany). The samples were injected in split mode using an autosampler AOC-20s / I (Shimadzu Deutschland GmbH). GC solution Chromatography Data System (Shimadzu Deutschland GmbH) was used for data processing. For the quantification of SCFAs, an external standard substance (Supelco TM WSFA-1 Mix, Supelco Sigma-Aldrich Co. Bellefonte PA) was employed.

[0026] The results are shown in Figure 1.

[0027] In mice fed a high-fat diet (HFD), the concentration of butyrate (butyric acid) in feces significantly increased 8 weeks after HFD intake. Such an increase was antagonized by the supplementation of SF68 at both 4 + 4 weeks and 8 weeks of treatment.

[0028] Example 2 Efficacy test on chicken weight gain The cage numbers of the chickens in the five treatment groups were randomly assigned to 12 replicates, resulting in a total of 60 test cages.

Table 2

[0029] Each cage was equipped with a nipple drinker and a feed tube with a capacity of 20.4 kg. The dimensions of each cage were 5'x4', and the stocking density per bird when raising 30 broilers per cage was 622 cm2 It was

[0030] The animals started being reared with fresh wood shavings. On the 5th day, all 60 cages were given materials to make 226 g of used bedding obtained from chickens that had not experienced diet DFM or enzymes.

[0031] A total of 1,800 one-day-old Ross 308 AP pure-line chicks (30 chicks / cage) were used.

[0032] On the first day of the test, the weights of 200 randomly captured chicks were measured and the average weight was obtained. Subsequently, the chicks were distributed to the cages in order from cage 1 (Table 2), and 30 chicks were randomly taken out of the wooden box, and the weights of the groups were measured so that the 30 chicks in all 60 groups were within ±10% of the obtained average weight.

[0033] During the entire test period, the chicks were illuminated. During the test period, all cages were checked at least once a day to manage the availability of feed and water and the temperature.

[0034] Each feed type (starter, grower, finisher) was prepared from one large basal feed formulated for each diet group and each treatment group to keep the sodium concentration as equal as possible across the treatment groups.

[0035] The meals were given according to the age of the chicks (starter from 0 to 14 days old, grower from 15 to 28 days old, finisher from 29 to 42 days old). The feed formulations are shown in Figure 2a (starter), 2b (grower), 2c (finisher).

[0036] All feeds were mixed on-site, and the amount of each feed type allocated to one chick was expected to be a maximum of 567 g for the starter, 1590 g for the grower, and 2270 g for the finisher.

[0037] The description of the supplementary feeds provided to the five test groups is reported in Table 3 below.

Table 3

[0038] All broilers had their body weight measured cage - by - cage at 0, 14, 28, and 42 days of age, and the feed weight was re - measured at 14, 28, and 42 days of age.

[0039] During the test period, the body weight, feed intake of broilers at 0, 14, 28, and 42 days of age, and the related feed conversion value (FCV) were measured. The results obtained are reported in the following table.

Table 4

Table 5

Table 6

Table 7

[0040] From the 28th day, a significant difference was observed between the treatment group and the control group. At the end of the test (the 42nd day), the combination of probiotics and butyrate numerically increased the average body weight compared to the butyrate administration group. (Figure 1) JPEG2025519442000009.jpg154149 (Figure 2) JPEG2025519442000010.jpg207140JPEG2025519442000011.jpg175140 (Figure 2b) JPEG2025519442000012.jpg205140JPEG2025519442000013.jpg203140 (Figure 2c) JPEG2025519442000014.jpg205140JPEG2025519442000015.jpg171135

Claims

**Claim 1** Use of Enterococcus faecium in combination with butyric acid or a salt thereof as a feed supplement for improving the health of intestinal cells and as a result improving the well-being of livestock and pets. **Claim 2** Use according to claim 1, wherein the Enterococcus faecium is a strain with deposit number NCIMB 10415. **Claim 3** Use according to claim 1, wherein the butyric acid or a salt thereof is sodium butyrate. **Claim 4** Use according to any of the preceding claims, wherein the Enterococcus faecium is in a microencapsulated form. **Claim 5** Use according to any of the preceding claims, wherein the butyric acid or a salt thereof is in a microencapsulated form. **Claim 6** Use according to any of the preceding claims, wherein the livestock are fattening turkeys, chickens, laying hens, lactating and weaning sows and piglets, fattening pigs, dairy cows, calves, and the pets are dogs and cats. **Claim 7** A feed supplement containing Enterococcus faecium in combination with butyric acid or a salt thereof for improving the health of intestinal cells and as a result improving the well-being of livestock and pets. **Claim 8** The feed supplement according to claim 7, wherein the Enterococcus faecium is a strain with deposit number NCIMB 10415. **Claim 9** The feed supplement according to claim 7, wherein the butyric acid or a salt thereof is sodium butyrate. **Claim 10** The feed supplement according to any one of claims 7 to 9, wherein the Enterococcus faecium is in a microencapsulated form. **Claim 11** The feed supplement according to any one of claims 7 to 10, wherein the butyric acid or a salt thereof is in a microencapsulated form. **Claim 12** The feed supplement according to any one of claims 7 to 11, wherein the livestock are fattening turkeys, chickens, laying hens, lactating and weaning sows and piglets, fattening pigs, dairy cows, calves, and the pets are dogs and cats.

Images