SE510497C2 - Additive for non-ruminant animal or human feed - Google Patents

Abstract

An additive (I) for a non-ruminant animal feed comprises \- 3 microorganisms selected from: (a) Pediococcus pentosaceus, (b) P. acidilactici, (c) Picia farinosa, (d) Dekkera bruxellensis, (e) Bacilli, (f) Streptococci, and (g) Staphylococci. (I) preferably also contain enzymes, organic acids and bacteriocins. The feed additionally contains \- 1 hydrolytic enzyme selected from (h) proteolytic enzymes (including those with trypsin-like and peptidase-like activities); (i) carbohydrate-splitting enzymes (including those with amylase-like and cellulase-like activities); lipolytic enzymes (including those with triacylglycerolase-like activities), peroxidase enzymes (including those with catalase-like activity) and transferase enzymes (including those with acyltransferase-like activities). (I) also includes \- 1 (preferably all) of lactic, acetic and succinic acids or their equivalents. It also includes \- 2 of the bactericins Pediocin A, Nisin, Pediocin AcH and P. acidilactice PAC 1.0.

Description

510 497 2 produces, is washed away and the onset of effect, when administered to animals, is either a zero level or a delay due to a) poor or no colonization, b) the destruction of microorganisms in the acidic part of the stomach, c) the production of enzymes and organic acids are delayed, which means that it is too late to have growth-promoting effects, d) no production of the essential lactic acids in live microbes. There is therefore a definite lack of efficiency.

Pathogenic bacterial infections of the stomach and intestines of animals such as pigs, and also of chickens, poultry and shrimp, are a major problem for both breeders and consumers. Although the breeder manages to maintain a very high hygienic standard in the breeding area, it is very difficult, and even impossible, to avoid such infections and during the weaning period for piglets, the development of severe diarrhea depends on immunological factors in the relationship between natural E. coli and lactic acid in the intestine. The frequent use of antibiotics to fight infections has become a major problem, due to the development of resistant strains of pathogenic bacteria. Perhaps the biggest problem is resistant strains of salmonella. Several of these bacteria have become resistant to many antibiotics, with the result that the animals die despite antibiotic treatment. Once the pathogenic bacteria have caused an imbalance, the animals become unable to gain weight in the normal way due to their reduced ability to digest the feed. In addition to the obvious financial losses for breeders, the quality of the meat often deteriorates and there is an obvious risk that those who consume the meat can become infected.

One group of animals that are very susceptible to bacterial infections are piglets that are often infected by the sow. When they are taken from the sow at the weaning period and the beginning of the intake of solid feed, they often develop serious intestinal problems. The problems are often so serious that some of the piglets will die.

The infections also cause major financial problems for the breeders, not only because some of the piglets will die but also because the breeding time of the infected animals will be unnecessarily long. The degree of severity of these problems with sick piglets varies from country to country. In Europe, about 15-3096 of the animals become infected, but in tropical countries the numbers are significantly higher, e.g. up to 40%.

Breeding broilers is another group with very big problems.

Infections of salmonella and carnpylobacter in the birds are very frequent and cause great economic damage to the breeder and can cause fatal infections in the consumers of the chickens. Salmonella infections in broilers and laying hens enter an uncontrollable stage worldwide as the salmonella bacterium develops a high degree of resistance to all types of antibiotics.

Once an animal has been infected with pathogenic bacteria, it is difficult, or impossible, to cure the animal with the antibiotics and probiotics that are currently available on the market. Shrimp farming is also facing very big problems, due to bacterially infected water, deteriorating the bottom layer of the ponds caused by uneaten feed, excrement and chemicals. All these environmental and pollution factors increase the risk of pathogenic infections in shrimp, leading to very large economic damage to growers. One of the most common pathogenic bacteria, which causes up to 100% mortality, is different from vibrio bacteria.

From an environmental and pollution point of view, the rearing of chickens, laying hens and pigs involves very big problems and there is an urgent need to reduce ammonia emissions.

Theoretically, the best way to solve the above-mentioned problems of growth-promoting infections without antibiotics and improved ecology would be to find a way to prevent the pathogenic bacteria from adhering to the mucosal surfaces or, for animals that have already suffered from an imbalance in the bacterial organism, to find a way to restore a normal bacterial ra ora that leads to lower mortality rates in animals and increased growth by using a composition of a natural "cocktail" with multiple symbiotic effects, consisting of living microorganisms, its inherent digestive enzymes, its inherent organic acids and its inherent bacteriocins, with a rapid onset and thereby strengthening of the immune system. Furthermore, the product would consist of the enzymes, organic acids and bacteriocins, which are produced by the living microorganism during the fermentation process. The rapid onset of the multiple symbiotic effects of the enzymes, organic acids and bacteriocins, which are metabolite substances from living micro-organisms, is necessary for a probiotic to be considered effective.

Summary of the Invention In a first aspect, the present invention relates to an animal feed additive or composition, characterized in that it comprises at least three and preferably seven of the following living microorganisms: a) Pediococcus pentasaceus b) Pediococcus acidilactici c) Pícia farinosa d) Dela / tera bruxelleltsis e) Bacilli f) Streptococcus g) Staphylococcus The invention is characterized in that it comprises enzymes, "digestive enzymes", which exhibit a variety of hydrolytic activities according to the following h) Proteolytic enzymes, e.g. trypsin- nch peptidase-like activities i) Carbohydrate-cleaving enzymes, e.g. amylase- and cellulase-like j) Lipolytic enzymes, e.g. triacylglycerol-lilining activities k) Peroxidase enzymes, e.g. catalase-like activities l) Transferase enzymes, e.g. acyltransferase-like activities and characterized in that it comprises at least three of the following organic acids: m) Lactic acid n) Acetic acid o) Amber acid and which is characterized in that it comprises the following bacteriocins: q) Pediocin A, Nisin, Pediocin AcH, P. acidilactice PAC 1.0 possibly together with animal feed, which may be known as such. The mixture of microorganisms will sometimes be referred to here as a "storage culture". A preferred animal feed according to the invention comprises about 8 to 15% water, about 15 to 30% total protein, about 0.5 to 5% total lipids, about 5 to 20% fiber, about 8 to 20% pf ash (minerals), about 30%. to 50% soluble, nitrogen-free material, about 6x10l to 3x10 ° living Pediococcus pentasaceus / g, from about 1x10 "to 1x107 living Pediococcus acidilcictici / g, from about 2x10" to 1x103 Picia farinosa / g, from about 21x10 "to 21:10 "Delzkera bruxellertsis / g, from about 2x10" to 4x108 Streptococcus / g, from about 25x10: to 6x107 Bacilli / g, from about 6x10 * to Gx107 Staphylococci, from about 0.2% to 5% lactic acid, from about 0.1 % to 3% acetic acid, from about 0.01% to 2% succinic acid and it preferably has an energy content of about 2-15 MJ / kg dry weight.

The above-mentioned microorganisms, enzymes, organic acids and bacteriocins preferably have i.a. the following properties: 1) Resistance to stomach acids 2) Heat resistance up to a maximum of 70 ° C 3) Ability to be pelletized 4) Stability in fresh and salt water 5) Non-toxic to animals and humans 6) Prevent infections' 7) Fight infections 8) Growth-promoting 9) Good colonization 10) Bacteriocidal effects' 11) Immunosuppressive effects.

According to the invention, there is provided a novel animal feed composition which has an unexpected effect both on animals having a normal bacterial och ora and, perhaps more importantly, on animals having a pathological bacterial fl ora.

The. The novel animal feed grain position of the invention, which will be described in more detail below, includes as essential components pediococcus bacteria, bacilli, streptococci and staphylococci, yeast cells (fungi), hydrolytic or digestive enzymes, organic acids and bacteriocins.

The mechanism by which the novel animal feed composition of the invention acts in the animals is not fully known, but we currently assume that the living microorganisms, according to the invention, compete with the pathogenic bacteria to restore normal balance and protect the normal bacterium. the competitive exclusion concept.

The organic acids will reduce the pH of the crayfish (chickens and laying hens) and the large and small intestine and make it difficult for the pathogenic bacteria to grow. Lactic acid will also kill the pathogenic bacteria through light. Both the living microorganisms and the lactic acid will, through their symbiotic action, stimulate the immune system when used in both animals and humans.

In the demand (for chicken), stomach and intestine, the digestive enzymes already present in the animal feed composition will act on the feed and break it down into more readily absorbable proteins, fragments of proteins and amino acids, whereby the total utilization of the energy-containing and other nutritional components of animal feed are improved. In any case, the result is that the animals, which have bacterial problems, can restore a normal (gastro) -intestinal balance. An additional effect is that the digestive uptake from the intestinal tract will increase. An extremely important result is that the animals' immune systems will be significantly improved and can effectively fight ongoing infections and / or infectious invaders.

An effect of feeding the animals with the composition according to the invention is that it restores / maintains a normal bacterial ear without causing any other problems. Another effect is that the composition has the ability to prevent infections and at the same time maintain a high level of healthy bacteria in the intestine without them leading to other problems. Another effect of the lyre focal composition according to the invention is that it has the ability to remove bacteria and also to improve the overall utilization of the energy-containing and other nutrient components in the animal feed. The animal feed composition defined above can be successfully used in conjunction with virtually any conventional animal feed and it will not be destroyed by the acidic conditions in the stomach.

The enzymes the organic acids and bacteriocins already present in the composition of the invention increase the rate of colonization of the living microorganisms so that they can continue to produce more enzymes, organic acids and bacteriocins, which are necessary for rapid replication of the living microbes.

The bacteriocins produced in the living microorganism have at least two different modes of action: a) they are produced to protect the living microbes against other bacteria that can kill them and b) they themselves have an antibacterial effect.

The stock culture and animal feed composition of the invention, its use and beneficial effects will be further illustrated in the following non-limiting Examples.

Example 1 - Preparation of animal feed Preparation of "feed culture" 1. 12 kg of soil, containing the microorganisms described above, are mixed with 30 kg of rice bran, 1 kg of soybean powder and 14 liters of water. The mixture is heated to 50 ° C for 12 hours. the temperature to 35 ° C and the mixture is kept at this temperature for 48 hours. The "stock culture" is dried to about 6% and is ready to use for the preparation of the animal feed composition.

Preparation of the animal feed composition 1. 500 kg of rice bran is mixed with 1.4 kg of the above-described stock culture. Water is added to a moisture content of 35%. 2. The mixture is spread in a layer of 10-15 cm and covered with plastic film to prevent water evaporation. The fermentation is complete after 8 days. 3. The plastic film is removed and the product is dried to about 7-9%. The product is then ready for direct use or for packaging for subsequent use, e.g. in paper bags. anal; s Water content 8.1% Total protein 22.796 Total lipid 3.1 ° / ¿> Fiber 11.7% Ash (minerals) 14.6% Soluble nitrogen-free substances 39.894 »211105 - 3x109 11.4 NIJ / kg weight Number of living microorganisms Energy content Example on the use of the product 1. Piglets - prevention of weaning diarrhea and other infections The aim of the study was to investigate 1% of the composition according to the formula mixed with normal feed and start at the weaning day (day 24) and end the experiment when the piglets had an average weight of 20 kg.

Results: Control Test group Number of piglets 188 178 Average age at start 24.04 days 22.24 days Average weight at start 6.78 kg 6.15 kg joint weight at end 15.60 kg 15.07 kg Average age at start 64.04 days 58, 24 days Mean trial period 40.00 days 36.00 days iw -uj 10 20 30 510 497 "8 Control Test group Mean growth increase 8.82 kg 8.92 kg Mean growth rate / day 0.221 kg 0.248 kg Salmonella infected day: 35 40 Number of infected suckling pigs' 73 52 Treatment with Norfloxazine injection day: 36 No Nor fl oxazine and Neomycin in the feed from day: 37 No Mortality, number of piglets' 42 (58%) 5 (1%) Conclusion: The trial could not be completed completely according to the protocol due to the salmonella infection .

The '3 infants in the control group, who developed salmonella infection, were very ill and 42 died within a few days despite antibiotic treatment. On day 64, the veterinarian decided to sacrifice all 188 piglets.

The piglets in the test group with the composition according to the invention had delayed infection compared to the piglets in the control group. The severity of the infection and diarrhea was small and only 5 died in the test group without the use of any form of injection of antibiotics or antibiotics mixed into the feed.

The composition also showed clearly better results in preventing salmonella infection and destroying salmonella bacteria compared to the traditional use of antibiotics. 2. Piglets - prevention of infection The aim of the study was to investigate the usefulness of piglets of 0.5% (5 kg of the composition / ton of feed) in mixture with normal feed and start day 24. The experiment was discontinued when the piglets gained a weight of 15 kg .

Results: Control Test group Number of piglets 143 133 Total weight 975 kg 954 kg Average weight at weaning 6.82 kg 7.17 kg 10 15 20 30 51715 4917 9 Control Test group Average age at weaning 25.42 days 24.08 days Total weight at the end 2 131, 5 kg 1961 kg Total number of piglets at the end 128 130 Mortality, number of piglets 15 (10.4996) 3 (2.26%) Weight at the end 16.65 kg 15.08 kg Wedel age at the end 66.19 days 60.78 days Mean trial period 40.77 days 36.76 days Mean growth 9.83 kg 7.91 kg Mean growth / day 0.245 kg 0.215 kg ADG 241.06 g 215.76 g FCR 1.56 1.77 FC / kg 14.05 15.95 Conclusion: l - The control group had gained more weight, but this was mainly due to the fact that they were 4 days older and that the growth rate of the piglets is much greater during these 4 days.

The frequency of weaning diarrhea in the control group is relatively high.

In the test group, no diarrhea was registered at all.

Mortality is significantly higher in the control group than in the test group. The high mortality was due to severe E. coli infection in the control group that occurred after the weaning period was over. The E. coli infection in the test group was very mild. 0.5? / <> - the dose of the composition is too low, but sufficient to protect against infections, but not sufficient to increase the growth rate. Sows The purpose of the study was to investigate the usefulness of 1% of the composition according to the invention in mixture with the normal feed given to the sows 4 weeks before farrowing and until weaning.

Result: Control Number of sows 54 Test group O! * J 10 20 25 30 510 497 Number of piglets Stillbirths Piglets Number of "Mummies" Small and defective piglets Total number of defective piglets Good live piglets Average birth weight Number of piglets per litter Breastfeeding sows Mortality at weaning Weaning weaning Weaning weaning / sow Conclusion: The weight birth weight was 30 g (2 teats were 9.7 9% in the control flu, comparing the difference of 51%.

The infants in the test group had a mean weight g (3.1%) greater than for the control group, which is the weaning period.

The reason for the higher mortality is weaker piglets and severity between 4-7 days after day 28. When the sows go on the composition 4 weeks before and 4 weeks after the pig, the composition increases the immune system, as is clear from the thicker milk and the infants' protection against infection. 10 554 Control 21 (3.79%) 5 (0.90%) 7 (1269/0) 33 (5.9G%) 521 (94.04%) 1.45 kg 9.65 51 51 (9.799f6) 470 7.04 kg 27.57 days 9.22 567 Test group 13 (2.29%) 10 (1.7G%) 12 (2.12%) 35 (6.17%) 532 (93.83%) 1 , 48 kg 9.33 55 26 (4.899 ~ ß) 506 7.26 kg 27.59 days 9.20%) higher for the sows in test group one. Mortality with 4.89% in the test group, a t at weaning, which was 220 r very important when entering the control group is, according to the veterinarian, more diarrhea. They were also ready for reproduction 4. Sows 'health and mortality before weaning The aim of the study was to investigate the use (60 g / day / sow) mixed with the normal feed compared to the sows' health and mortality before weaning. of 1096 of composi- 10 20 25 30 510 497 11 Results: Pig Control Test group Number of sows 10 10 Total live born piglets 102 95 Average birth weight 1.29 kg 1.48 kg Average total number of piglets / litter 11.1 10.0 Average number of pigs born / litter 10.2 9.5 Average number of stillborn pigs / litter 0.5 0.2% stillborn piglets 4.5 2.0 Average number of "mummies" / litter 0.4 0.3 96 "mummies" 3.6 3.0 Weaning Number of weaned sows 10 10 Total weaned piglets 86.0 94.0 Weaned piglets / litter 8.6 9.4 lVI mortality before weaning 15.7% 1.1% Mean weight at weaning 6.05 kg 6.33 kg Average age at weaning 21.4 days 21.8 days Conclusion: The mean weight in the test group at farrowing is 190 g or 15% higher than in the control group, which must be considered significant.

Mortality before weaning of 15.7% in the control group is due to severe weaning diarrhea. The mortality in the test group before weaning of 1.1% must be assessed as a direct result of the composition according to the invention, which keeps the sows very healthy and the milk contains more immunoglobulins which are essential for the piglets and can protect them against infections.

The difference in mean weight at weaning of 280 g, or 5%, is also significant. 5. Pigs - blood analysis and general analysis values The test was performed by Japan Food Research Laboratories in collaboration with Yamaguchi University Agriculture Department.

A summary of the test is that the composition according to the invention has a favorable effect as follows: Meat (red muscle / fat 85/15%) Control animals Test group Water content 63.4% 64.1% Protein 18 .5% 19.5% Fat 1G .8% 15.1% Cholesterol 50.0 mg 44.4 mg pH 5.7 5.8 Conclusion: Breeding pigs for slaughter with the composition according to the invention during the last 3 months reduces the levels of cholesterol and fat significantly, which. must be considered very favorable to consumers. The reason for this is the conjugated fl all-acid enzymes, which have been produced during the preparation of the composition, and also produced from the living microbes in the composition when given to pigs. See Tables 1 and 2, Figs. 1-5. 6. Slaughter pigs - Prevention of diarrhea and shorter rearing time The study was organized by Prozyme AB, Uppsala, Sweden in collaboration with the University of Agriculture, Department of Animal Breeding and Genetics; Animal health services, Uppsala, Sweden and Farmek slaughterhouse, Uppsala, Sweden.

The test group and the control group included 27 animals each. The test group had an initial mean weight of 32.6 kg and the control group had a mean value of 32.2 kg.

The pigs in each group were given the same amount of feed, but in the test group the animal feed contained 0.7 5% of the feed additive according to the invention. All animals were weighed once a week.

Results - Test group, 27 animals: A total of 5950 kg of feed was needed to reach final body weight, ie. one. average value of 220.4 kg of feed per pig. The first pig was slaughtered on day 62 and the last day 154. A total of 2546 days were needed to reach the final body weight, ie. a mean value of 94.3 days / pig. FCR 3.03 - see Table 1 below. None of the animals in this group needed medical treatment during the study. No side effects or abnormal behavior were observed in the test group in 51 Ü 497 13 Control group, 27 animals: A total of 7335 kg of feed was needed to reach the final body weight, ie. on average 271.7 kg of feed per pig. The first pig was slaughtered on day 76 and the last pig on day 154. A total of 3043 days were needed to reach the final body weight, ie. an average of 112.7 days / pig. FCR 3.76, see Table 1 below. Seven animals in this group needed medical treatment due to stomach disorders.

Data: Total number of animals Body weight at start Body weight at end Total amount of feed Feed, mean value per pig Total number of days Days, mean value per pig FCR End date / number of pigs / - final body weight Table 1 Test group 27 32.6 + 13 / -1G kg 105, 2 + 8 / -3 kg 5950 kg 220.4 kg 2546 days 94.3 days 3.03 62/3 / 103-105 76/6 / 103-110 84/2 / 104-105 91/5 / 103-106 98/2 / 103-106 105/4 / 103-109 112/1/103 147/1/104 154/1/105 1. Broilers - Prevention of salmonella The study was carried out by Spelderholt DLO Institute for Animal Science and Health Agriculture Research Department DLO-NL PO Box 15 7360 AA Beek- Control group 27 32.3 + 11 / -17 kg 104.5 + 5 / -6 kg 7335 kg 271.7 kg 3043 days 112.7 days 3.76 76/2 / 103-108 84 / 3 / 104-105 91/2 / 104-105 93/5 / 103-108 105/2 / 103-105 112/2 / 103-104 119/1/105 126/3 / 105-109 133/1/104 147/3 / 98-107 154/3 / 105-106 10 15 20 30 510 497 ~ i "ttï7 Ä 14 mountains, the Netherlands.

Salmonella challenge in broilers Broilers in 8 cages were individually infected orally on day 14 with 10 'cfu of Salmonella infantis Nal and in the other groups 5 were marked per cage, simulating the natural infection. Oral infection with 5x107 cfu of Salmonella infantis Nal day 14. Salmonella status of the experimental chickens before infection was tested by stool samples. At the time of sampling (1 week after infection), 5 broilers were slaughtered from each cage and in caeca the number of salmonella was estimated.

Feed The feed included 58 mg / kg Zinc bacitracin as a growth promoter and 3 mg / kg halofuginone as a coccidiostatic. The feed was prepared for the entire trial period and administered "ad libituni". The feed additive according to the invention was incorporated into the feed for pelleting (1%, weight). The pelletizing temperature was 70 ° C.

Microbiology 7 days after infection, 6 chicks were randomly selected from each litter (48 chickens per group) and slaughtered. Caeca was then taken out and after sterilization of the surface a sample of the contents was taken aseptically. The cfu count of Salmonella in / arzlis Nal was estimated by plating on "Brilliant Green" agar (Oxoid CM 329) with 20 ppm Nalidex acid. Ten-fold dilutions of the caecal content of Buffrat Pepton-Water (Oxoid CM 509) were incubated and plated on BGA agar after enrichment.

Statistics The results were analyzed with the student's t-test.

Results and discussion Infection with very high doses of salmonella (5x1O7) was established as expected and after a few days the number of salmonella-positive chickens was at an acceptable level of about 70%. The mean number of salmonella cfu in the caecal content of broilers from both test groups was significantly lower than in the control group (P <0.01). 10 15 20 30 051004970 '15 Table 1 shows results from individual cfu bills in the caecal content. In the table, the number of broilers with a certain level of salmonella cfu has been marked. From these results one can calculate Infection Factors (IF) and Protection Factor (PF).

IF is the geometric mean of the number of salmonella per g caecal content of all chickens in a particular group. From this IF, pF can be obtained, by dividing the IF value in the control groups by the same for the treated group.

Table 1. Number of chickens with a certain number of Salmonella cfu per gram caecal-lialt, IF (infection factor) and PF (protection factor) Week 1 - Orally infected Number of salmonella per g caecal content no 10 102 10 "10 * 10 * 106 10" ' > 1o3 IF control 40 0 0 1 G 10 12 9 1 1 4,7 Test 40 10 3 0 1 7 6 4 2 3,6 PF: 1,3 -J Week 1 - via offspring.

Number of salmonella per g caecal content n 0 10 102 10 * 10 '10 "' 10" * 10 *> 10S IF 1 <0nn ~ 0114o 2 0 0 3 3 14 11 2 5 5.3 Test 40 20 0 2- 1 2 0 e 1 1 2.5 PF: 2.1 At the sampling date (1 week pi), salmonella was missing in a larger number of the broilers from the test group than from the control group (Table 2).

Table 2. Salmonella analysis of caecal samples of salmonella-negative chickens. n = 40 oral offspring control test control test Week 1 0 10 1 20 Conclusion: Administration of the composition according to the invention to broilers contributes significantly to early exclusion of salmonella. The rapid onset of action is due to the multiple symbiotic action of the living microorganisms, enzymes, organic acids and bacteriocins that make up the composition of the composition. The results clearly show the effectiveness of the composition in removing salmonella infections from chickens. 8. Broilers - Growth increase The aim of the study was to compare the composition according to the invention, using 1% mixed with feed materials that do not contain any antibiotics, with a control group with normal feed that contains' antibiotics, for breeding broilers, one day old chickens for slaughter. The study was conducted by Chaiyaree Farm Co., Ltd. Panat Nikhom, Thailand.

Results Control Test group Number of chickens 10,200 10,200 Age of chickens 1 day old 1 day old Final age 46 days 46 days Drugs used during the experiments: - Nlintervít day 1 day 1 - IB A 3 day 2 day 2 - Tylan day 3 day 2 - Permasol day 6 day 9 - ND clone day 10 day 8 - IBD Blen day 14 day 13 - Tylosin SP day 17 None - Post-vaccine day 18 day 15 - Chlorine-Ery day 28 day 28 - Triane day 40 day 40 Dead chickens: Week 1 82 pcs 57 pcs Week 2 115 pcs 106 pcs Week 3 69 pcs 50 pcs 10 15 30 Week 4 Week 5 Week 6 Week 7 Total: 33 pcs 43 pcs 118 pcs. 325 pcs 785 pcs Sick and weak chickens that are removed.

Week 1 Week 2 Week 3 Week 4 Week 5 Week 6 Week 7 Total: Control 10 pcs 24 pcs 2G pcs 18 pcs 16 pcs 26 pcs 111 pcs 231 pcs 516149? 17 27 pcs 33 pcs 42 pcs 45 pcs 360 pcs Test group 26 pcs 20 pcs 6 pcs 0 pcs 0 pcs 1 pc 89 pcs 142 pcs Total dead and sick chickens, number: Week 1 Week 2 Week 3 Week 4 Week 5 Week 6 Week 7 Total: 92 (0.90%) 139 (1.36%) 95 (0.93 ° / í> 51 (0.50% 59 (0.58%) 144 (1.41 ° / í> 436 (4 , 2'T%) 83 (0.81 "/ 6) 126 (1,249 ~ 6) 56 (0,55f: / 0) 27 (ozeeß) 33 (osm) 43 (oazet) 134 (1,3196) 1016 ( 9.96'5 ~ í,) 502 (4.92%) Live chickens 9184 Total kg: 176985131 Meclelvíkt 1.93 kg Total feed consumption, kg 36.195 FCR 2.05 Conclusion: The experiment as such is very well controlled by Chaiyaree Farm Co. , Ltd. 9698 1984696 2.05 kg 40,489 2.04 10 15 20 30 510 497 18 The chickens The last antibiotics 6 days before slaughter and in the control group The chickens also had diarrhea and lost weight and had a high total mortality.

During the last week, when antibiotics are banned, the chickens lost weight and mortality increased, which is a very big problem for the breeders to overcome. During the last week, there is a risk of re-infection of Salmonella and Campylobactei ', which can spoil the breeders' entire business.

The differences in mortality are significant, as are the heavier chickens in the test group.

The difference of 120 g / chicken in combination with more live chickens is extremely large from an economic point of view. 9. Broiler - Cost / Revenue Study Background: SUNEK FOOD LTD., Saraburi, Thailand is a company that exports broilers to Japan. They are among the 10 largest companies in Thailand in this area.

Every company that exports broilers must follow very strict rules regarding residues of antibiotics, chemicals and any pathogenic bacteria present in the meat.

This company has a production capacity of 60,000 broilers / day, or about 19 million broilers / year. This means that they need to 'perform the broiler breeding in a very professional way to maximize the profits and establish a good relationship with their Japanese customers.

SUNEK has extensive experience of tests with "probiotics" as an alternative to antibiotics to promote growth and also to rule out salmonella contamination of the meat. To date, they have not found any type of probiotic available on the market that meets all the criteria.

We can describe the experimental conditions using the composition according to the invention as follows: Criteria: 10,000 day-old chickens in each group were placed in 2 different houses, about 50 meters apart.

The control group and the test group would have the same type of antibiotic-free feed and the same standard medical treatment regimen. The same number of workers would take care of the two groups. The chickens in the test group would be given 0.55% (5 kg / 1 ton feed pellets) of the composition in mixture with feed pellets. Slaughter was to take place on days 44-46 at its own slaughterhouse industry in the province of Saraburi.

Results: Test group Control group Number of day-old chicks: 10,600 10,000 Mortality: Test group Control group Week 1,119 (1.12%) 180 (1.80%) Week 2,131 (12,496) 203 (2.03%) Week 3,112 (1.06 %) 172 (1.72%) Week 4 115 (1.09'3f¿) 138 (1.38%) Week 5 97 (0.92%) 211 (2.1l% week s 171013194.) 1so (1 , p6 Week 7 134 (1.26%) 78 (0.78%) Total: 879 (8.29 ° / ó) 1168 (11.68% Anni: Week 7 a total of 4 days for the test group and in the control group 3 days.

Feed consumption: Week 1 1,500 kg 1,350 kg Week 2 3,060 kg 2,850 kg Week 3 5,070 kg 4,770 kg Week 4 7,140 kg 6,540 kg Week 5 7,950 kg 7,830 kg Week 6 9,570 kg 9,930 kg Week 7 4,080 kg 2,160 kg Total: 38,370 kg 35,430 kg Note: Week 7 for the test group was total. 4 days and in the control group 3 days.

IVIedical program: Test group p Control group Day 1 IB vaccine IB vaccine Day 1-3 Tylosin Tylosin 10 20 510 497 Day 10 Day 14 Day 15 Day 22 Day 29 Day 35 Total body weight: Number of chickens for slaughter: Total feed consumption: FOR: half weight / chicken: Cost / revenue analysis: Costs: Day-old chickens .Drugs Feed Testp rcver Work TOTAL Income: TOTAL: Total net income: Net income / chicken: Discussion: 20 ND vaccine IBD vaccine Amcolístíne oxramine Tetramycin 7. 21 kg 2.05 1.92 kg Test group USD 4,240 G36 USD 9,669 USD 191.84 USD 212 USD 14,834, G8 USD 20,196 USD 5.2G1.32 USD 0.496 USD ND vaccine IBD vaccine Amcolistine Tetramycin Tetramycin Noroxacin 16,000 kg Control group 8,832 pcs 35,430 kg 2.21 1.81 kg Control group 4,000 USD 600 USD 8,928 USD O USD 200 USD 13,728.36 USD 17,280 USD 3,551.64 USD 0.356 USD The experiment was 100% performed and controlled by SUNEK FOOD LTD.

Biofeeds staff have visited the farm once / week to collect data and inspect the health of the chickens. SUNEK decided to use the same standard medical program for both groups to obtain a comparative trial.

Conclusion: The differences between the two groups are, in all respects, significant.

The degree of mortality in the control group of 11.68%, compared with 8.29% in the test group, is about 29%, which is definitely statistically significant. The difference in FCR of 2.05 in the test group versus 2.21 in the control group is more than 7% and must also be considered significant. Most important, however, is the cost / benefit analysis, which clearly shows the benefits of using the composition of the invention and the differences could have been greater if the use of antibiotics after day 3 had been ruled out, as this type of antibiotic treatment is only prophylactic. However, the net income per chicken in the test group is 12.41 Baht / chicken and is in the control group 8.89 Baht / chicken, which is a difference of 40%.

The differences between the composition and other types of microbial products are very large, due to the fact that the composition can be classified as a ready-made "Biotic System", which initially contains all the necessary ingredients such as living microbes, organic acids, enzymes and bacterio- ciner This together is a guarantee that you get good early colonization in the crayfish and a direct effect of the enzymes on the fodder in the crayfish.

The organic acids will first of all lower the pH and make the situation unfavorable for pathogenic bacteria and these together with especially a) the milk acid which has an immediate bacteriostatic effect and b) the total immune-boosting effects of the living microbes are what is needed to keep the chickens in a healthy condition.

When using products that contain live microbes in chickens, it is very important that the beneficial effects start directly in the craving and that there is minimal damage to the live microbes in the acidic part of the stomach, as the chickens have poor digestion mechanism and low nutrient uptake.

The composition is effective from the first second, to be compared with traditional probiotics which act very slowly because they have to start the production of their own metabolites before they can function. The above mode of action is the reason for the excellent net income obtained when using the composition.

We can also add that the risk of salmonella and campylobacter infections in chickens is reduced when using the composition, which is a very big problem worldwide. Something that could also be registered was the reduced smell of ammonia from the test group, compared with the control group. 10. Shrimp and "Prawns" Mussels grown in ponds are very susceptible to bacterial infections. Once an infection occurs, all the shrimp or mussels in the pond die. To minimize the risk of infections, the composition according to the invention is used in a concentration of about 20 g / m2, which is spread evenly over the empty bottom of the pond. The composition works for 7-10 days and after this time the concentration of pathogenic microorganisms is minimized to a level that does not damage the shrimp / mussels later.

After storage of post larvae, the composition according to the invention is used twice a week with 2 kg / 1600 m2. In addition, 1% of fine powder (mesh size 60) of the composition is also mixed with feed tablets and given to the shrimp every day.

Conclusion: The prawns in the ponds, which have been treated with the composition, as well as feed additives, grow to harvest size within 100 days, compared with the control, which on average is 120 to 150 days. The yield is much higher and the protection against various infections such as tiex. Vibrio bacteria, viruses and "yellow head" and "VVhite Spot" viruses. 11. Shrimp Vibrio Infection Two ponds with faecal samples of Vibrio bacterial infection were taken from both ponds. The amount of Vibrio bacteria in the faeces after culture was 21-1106 cfu / g. In one of the ponds, the water was treated with 3 kg of the composition twice a week for 2 weeks. The feeding tablets were also mixed with the composition with 1 kg of the composition / 100 kg of feeding tablets. The other dam did not receive any treatment at all.

After 2 weeks, faeces from the shrimp were collected from both ponds and grown.

Result: The control pond without any treatment had 8x10 ° cfu vibrio / g and in the pond treated with the composition according to the invention there were 53x10 ”cfu 10 15 20 30 510 497 23 vibrio / g faeces. Cats and dogs A number of both sick and healthy cats, kittens and dogs have eaten the composition of the invention every day for more than G months. Animals with digestive problems have recovered within a few days and healthy animals are kept in good condition, with accompanying normal faeces, good fur condition and less odor from the animals. 13. Horses 50 horses to be trained for trotting horses and suffering from intestinal problems leading to loose stools, weight loss, malnutrition, poor coat and general condition were given 60 g / day of the feed additive according to the invention for 2 weeks. After 2 weeks, the dose was increased to 180 g / day. The feed additive was mixed with the normal feed to spread throughout the day.

Result: size. After 4 weeks the horses started to gain weight and the condition of the coat and Already after 3 days the faeces returned to normal color and the normal overall condition improved. At this time, the horses were able to resume training in a normal manner. 14. Improving the environment The smell from broilers, partridges and pig farms is a very big problem and also a pollution problem that results from intensive cultivation. When using the composition according to the invention as a feed supplement for more than G months, the odor from the cultures subsided, which can easily be detected by the reduced odor of ammonia. Even after 1 month of using the composition, the condition of the farmers changes dramatically. 15. Degradation of animal urine A 2 meter deep layer of sawdust was applied in a special container, 100 m long, G m wide and 5 m deep. 50 tons of urine were sprayed over the sawdust layer and 5 kg of the composition according to the invention was added per 40 cm 3. After 5 hours, the first 1-meter layer mechanically stirred up air sucked in through a brower. An odorless aqueous solution was gelled through the sawdust layer. This odorless water contains less than 100 ppm free nitrogen and hydrogen sulfide as pollutants. 50 tonnes of urine can be decomposed 10 | ..i (TI 20 30 510 497 24 per day for 18 months. After 18 months, the upper sawdust layer of 1 meter must be replaced and new sawdust added. This method of decomposing urine is excellent to avoid environmental problems.

Similarly, the additive of the invention can also be used to decompose digested sludge and the product obtained can also be used as a fertilizer and for composting. 16. Use in humans - Acute bacterial infections 10 persons suffering from acute intestinal problems, caused by bacterial infections, have been given the composition according to the invention. 3 g 3 times / day for 1-3 days.

Results: 6 of the people responded within 12 hours with a lack of pain and discomfort and no diarrhea. 4 people had a normal situation after 24 hours. 17. Patients with nutritional problems Cancer patients normally suffer from digestive problems and they cannot break down food in the normal way, which means that they receive a very poor nutritional supplement, which is a major problem for the immune system to function. Five patients admitted to hospital, suffering from cancers leading to severe weight loss and general poor condition, were given the composition of the invention in admixture with drinking water, 5 g / day.

Result: After 6 days, the problems with loose stools disappeared and the second week a small weight gain could be registered. The appetite returned and after 4 weeks the patients gained significant weight and the general state of health improved. 18. Comparative experiments with the composition according to the invention without enzymes, organic acids and bacteriocins.

The purpose of the following study was to investigate whether the enzymes, organic acids and bacteriocins are needed in the composition according to the invention. Broilers have been used for evaluation and use of growth rate and mortality as an analysis method. The fermented product according to the preparation method was washed with water and filtered through a very fine mesh to separate rice bran and living microorganisms from the enzymes, organic acids and bacteriocins. The residue was washed with water. 10 15 20 30 5 1 0 4 9 7 25 No detectable level of living organisms could be detected in the aqueous phase and no detectable level of organic acids and enzymes could be detected in the rice bran and the rest of living microorganisms. The fermented product according to the preparation method was washed with water and filtered through a net to separate the rice bran from the living microorganism, the enzymes, the organic acids and the bacteriocins. The following test was performed with day-old chicks and ended on day 45, with 1000 chickens in each group and the same estimated amount of micro-organisms previously used, clvs. 1% additive to normal feed, see Table 1: Table 1 Group Growth rate Mortality% average weight 1. Control (no additive according to the invention) 1.90 kg 10.75% 2. Composition according to the invention 2.10 kg 4.63% 3. Composition according to the invention (without enzymes, organic acids and bacteriocins) 1.94 kg 8.40 ° -6 4. Aqueous phase (only enzymes, organic acids and bacteriocins) 1.90 kg 9.10% 5. Aqueous phase (only microorganisms, without enzymes, organic acids and bacteriocins) 1.96 kg 8.50% G. Aqueous phase (containing live microorganisms, enzymes and organic acids and bacteriocins) 2.15 kg 4.15% Conclusion: The experiments were performed simultaneously on the same test farm. The number of chickens (1000 in each group) is large enough to assess the results.

The results clearly show that the combination of living microorganisms, enzymes, organic acids and bacteriocins according to the invention is needed for rapid initiation, which has led to an increased growth rate in test groups 2 and 6 compared to control group 1 and the other groups. Test group 6 showed slightly better results than group 2, which can be explained by the fact that, when the aqueous solution is mixed with the normal drinking water, all the chickens get the composition in a better and simpler way than when the composition is mixed with feed pellets.

Blood analysis of pigs follow-up with the composition for 3 months Table 2 Control group 920228 920307 92.031? 920406 920415 920424 929520 Table 3 Test group 920228 92.03.07 92.03.17 92.04.06 920415 92.04.24 920520 TG: Triacylglycerol TG mg / dl 76.6 28.7 43.6 20.2 84.0 35.1 35 .1 TG mg / dl 53.2 26.6 22.3 18.1 29.8 14.9 TC mg / dl 89.9 98.0 96.0 66.0 85.9 92.9 89.9 85 .9 70.7 80.8 71.7 62.1 TC: Total cholesterol PL: Phospholipids FFA: Free fatty acids GL: Glucose PL mg / dl 128.3 138.0 137.0 101.1 114.1 141.3 96.7 PL mg / dl 107.6 127.2 93.5 110.9 103.3 75.0 26 FFA umol / l 157 216 255 98 153 110 212 F FA urnol / l GL Ing / dl 130.9 99 .0 85.3 100.0 164.7 115.2 79.4 / 71 L] Ing / dl 138.7 106.4 107.4 164.7 139.2 74.0

Claims (16)

10 15 20 25 30 ål- PATENTKRAV 51th 497i ¥ í ¥ Functional foodstuff, characterized in that it contains at least one of the following micro-organisms: a) Pediococcus pentosaceus b) Pediococcus acidilactici, and at least one of the following micro-organisms: c) Picia farinosa d) Cover bruxellertsis, and at least one of the following micro-organisms: e) Bacilli f) Streptococcus and g) Staphylococcus, and at least one of the following hydrolytic enzymes: h) Proteolytic enzymes, i) Carbohydrate-cleaving enzymes, j) Lipolytic enzymes, k) Peroxidase enzymes and 1) Transferase enzymes, and at least one of the following organic acids: m) Lactic acid n) Acetic acid o) Succinic acid, and at least two of the following bacteriocins: q) Pediocin A, Nisin, Pediocin AcH, P. acidilactice PAC 1.0 Functional food according to Claim 1, characterized in that it contains all the living microorganisms a) - g). Functional food according to one of the preceding claims, characterized in that it contains all the enzymes h) - 1). Functional food according to one of the preceding claims, characterized in that it contains all the acids m) - o). 10 20 25 30 510 497 2.53 Functional food according to one of the preceding claims, characterized in that it contains all the bacteriocins q). Functional food according to claim 4, characterized in that said proteolytic enzymes include enzymes having trypsin- and peptidase-like activities, said carbohydrate-cleaving enzymes comprising amylase- and cellulase-like activities, said lipolytic enzymes comprising triacylglycerol-like activities , k) said peroxidase enzymes include catalase-like activities and 1) said transferase enzymes include acyltransferase-like activities. Functional food according to any one of the preceding claims, characterized in that it contains about 8 to 15% water, about 15 to 30% total protein, about 0.5 to 5% total lipids, about 5 to 20% fi brer, about 8 to 20% ash (minerals), about 30 to 50% soluble, nitrogen-free material, about 6x104 to 3x109 live Pediococcus pentosaceus per g, from about 1x103 to 1x107 live Pediococcus acidilactici per g, from about 2x103 to 108 Picia farínosa per g, from approx. 2x103 to 22:10 ”of Dekkera bruxellensis per g, from about 2x103 to 414108 Streptococcus per g, from about 8x102 to 6x107 Bacilli per g, from about 6x104 to 6x107 by Staphylococci, from about 0.2% to 5% lactic acid, from about 0.1% to 3% acetic acid, from about 0.01% to 2% succinic acid. Functional photo substance according to one of the preceding claims, characterized in that it has an energy content of approximately 2-15 MJ / kg, dry weight-based. Functional food according to any one of the preceding claims, characterized in that said microorganisms, enzymes, organic acids and bacteriocins have at least one, and preferably all, of the following properties: 1) Resistance to stomach acids, 2) Resistance to heat up to a maximum 70 ° C, 3) Ability to be pelleted, 4) Stability in fresh and salt water, 5) Non-toxic to animals and humans, 6) Prevents infections, 7) Cures infections, 10 15 20 25 s10t497ï 27 8) Growth promotion de , 9) Good colonization, 10) Bacteriocidal effects and 11) Immunosuppressive effects. Use of a functional food according to any one of the preceding claims for the manufacture of a preparation intended to maintain a normal bacterial fl ora and restore a pathological bacterium fl ora in a human. Use of a functional food according to any one of claims 1 to 9 for the preparation of a preparation intended to increase the digestive uptake from the intestinal tract. Use of a functional food according to any one of claims 1 to 9 for the preparation of a preparation intended to improve the immune system of a human. Use of a functional food according to any one of claims 1 to 9 for the preparation of a preparation intended to prevent weaning diarrhea and other infections. Use of a functional food according to any one of claims 1 to 9 for the preparation of a preparation intended to prevent or reduce salmonella and / or campylobacter. Use of a functional food according to any one of claims 1 to 9 for the preparation of a preparation intended to prevent acute bacterial infections in a human. Use of a functional food according to any one of claims 1 to 9 for the preparation of a preparation intended to reduce the amount of cholesterol and / or fat in a human.