WO2015193845A1

WO2015193845A1 - Acidophilic yoghurt

Info

Publication number: WO2015193845A1
Application number: PCT/IB2015/054617
Authority: WO
Inventors: Jiri SVIHALEK
Original assignee: Yoggies, S.R.O.
Priority date: 2014-06-19
Filing date: 2015-06-19
Publication date: 2015-12-23

Abstract

Acidophilic yoghurt made of a mix of cow's milk and ewe's milk containing live milk fermentation bacteria beneficial to alimentary tracts of dogs or cats with reduced content of lactose. The production process employs a newly developed mix of bacteria used for production of yoghurt and bacteria used for production of acidophilic milk. The live bacteria content of 8, 2.10⁷CFU/g and the lactose content of 1,3 % in the product is retained for the minimum period of 42 days when stored at the temperature of 4°C.

Description

Acidophilic Yoghurt Technical Field

Nutrition of animals, particularly cats and dogs

State of the Art

The current dry nutrition, e.g. granules, is produced with a large share of cereals. This may cause problems as the stomach and intestines are not capable of responding to such a strong stimulus (cereals) and do not produce enough digestive enzymes and acids. Therefore, the food may not be adequately digested due to insufficient condition of digestive intestinal microflora.

The fat contained in granules must be preserved to allow for long shelf life of the granules. This is provided for by preservatives, e.g. BHA, BHT, etc. These may cause deterioration of immunity, have adversary effect on production of erythrocytes, and some are suspected to cause cancer.

Inadequate digestion in the stomach and intestines due to poorly digested proteins means accumulation of water, rapid peristalsis of intestines and diarrhoea.

The patent document EP 1151675 B l describes a food supplement consisting of a carrier produced by milk fermentation from Lactobacillus bulgaricus and Streptococcus thermophilus, with subsequent permanent termination of fermentation by heating of the product. The carrier is coagulated fermented milk with non-living bacteria. Thanks to that, it will last long without the need for storage at low temperatures. The product may be stored at room temperature for over 3 months, potentially over 6 months. It only supplies the animal with milk proteins of undocumented composition and with unknown quantity of lactose, which is usually not taken well by dogs and causes digestion problems.

Milk is a source of proteins and calcium, but it is not suitable for dogs and cats. It contains lactose (milk sugar) for which the organisms of dogs and cats do not have adequate enzymes, i.e. they do not produce enough lactase in their alimentary tract to process milk. Undigested lactose then causes diarrhoea. Therefore, it is recommended to feed sucklings and adult animals only with specially produces delactosed milk intended for cats and dogs. Sucklings have an enzyme in their intestines that can digest lactose, but adults lose this ability.

The most important bacteria for the alimentary tract of humans and animals is Lactobacillus acidophilus. It is characterised by production of acid metabolites in high concentration, which aids in maintaining rather acid environment required for healthy alimentary tract.

Principle of the Invention

Remedy of the above deficiencies of today's nutrition is aided by a new acidophilic yoghurt made of a mix of cow's milk and ewe's milk, which contains live milk fermentation bacteria beneficial to alimentary tracts of dogs and cats with reduced content of lactose. The product is made using a newly developed mixture of bacteria used for production of acidophilic milk. It is a composite milk bacteria culture for production of Acidophilic Yoghurt containing a mix of Lactobacillus delbrueckii subsp. Bulgaricus YOG 2 - LB and Strepthococcus thermophilus YOG 2 - ST, Lactobacilus acidophilus YOG 1 - LA and Lactobacilus helveticus YOG 1 - LH. The benefit of this composite culture may be further enhanced by addition of another milk fermentation bacteria - Bifidobacterium animalis or Lactobacillus animalis or their mix.

The resulting product can be stored for the minimum of 42 days at storage temperature of 4°C, the content of live bacteria is 8,2.10⁷CFU/g and the content of lactose only 1.3 %. Samples of the product with this lactose content were administered to dogs, the findings being that the product had no negative effect on their digestion and they can digest all of the components of the acidophilic Yoghurt - live cultures supporting intestinal microflora especially of dogs and cats as well as nutritive components of milk, particularly ewe's milk. Dogs found acidophilic yoghurt very tasty and always consumed the entire portion.

Acidophilic yoghurt is produced by fermentation of a mix of ewe's milk and cow's milk; the proportion of ewe's milk should be 20% at minimum, while 50% is recommended. The proportion of ewe's milk is a significant factor in reduction of lactose content. If a composite culture with 50% proportion of ewe's milk was cultivated, the residual lactose quantity dropped to 38% after 7 days of storage, while with 70% proportion of ewe's milk, the residual lactose quantity was as low as 27%. The composite milk bacteria culture for production of acidophilic yoghurt contains bacteria deposited in CCM in accordance with the Budapest Treaty: mix of Lactobacillus delbrueckii subsp. bulgaricus and Strepthococcus thermophilus CCM 8532, Lactobacilus acidophilus CCM 8534 and Lactobacilus helveticus CCM 8533. The composite culture consists of four strains in two groups in 1 :2 ratio. Group 1 contains Lactobacilus acidophilus CCM 8534 and Lactobacilus helveticus CCM 8533 in 1 : 1 ratio, while Group 2 contains Lactobacillus delbrueckii subsp. bulgaricus and Strepthococcus thermophilus CCM 8532 in 1 : 1 ratio. The composite culture contains one part of Group 1 and two parts of Group 2.

The composite culture can benefit from addition of Group 3 made of Bifidobacterium animalis or Lactobacillus animalis or their mix; Group 3 should not constitute more than 20% of the entire content of the composite culture.

The cultivation is commenced by addition of at least 1% of the composite culture to the milk content; it is always necessary to adhere to the condition that the quantity of Group 2 culture is identical to the quantity of Group 1 cultures or up to twice as high. Cultivation proceeds for 10 to 15 hours at the temperature of 36 to 42°C. After that, the product can be stored in cold environment in order to ensure long storage period of 42 days at minimum. If possible, cultivation should take 12 hours at the temperature of 40°C.

The product maintains viability of all bacteria of the composite culture in the quantity of 10⁸ CFU/g for the period of two weeks and 10⁷ CFU/g for additional four weeks.

List of pictures in the drawing Fig. 1 :

Graphic representation of dependency of the content of lactose in the product on the storage period and the type and quantity of groups of bacterial cultures. Three types of cultivation environment were observed: Composite culture (YOG 2 - LB, YOG 2 - ST, YOG 1 - LA, YOG 1 - LH) injected in milk in the concentration of 1 %, 2,15 % and 3,25 %, with the following designation: 2+\_\ %, 2+l_2% and 2+l_3 %., Group 1 cultures injected in milk in the concentration of 1 %, 2,15 % and 3,25 % with the following designation: 1 1 %, 1 2% and 1 3 %, and Group 2 cultures injected in milk in the concentration of 1 %, 2, 15 % and 3,25 % with the following designation: 2_\ %, 2_2% and 2_3 %.

Fig. 2: Product lactose values during storage for the period of 42 days. Milk was injected with Group 1 cultures in the quantities of 1 %, 2, 15 % and 3,25 %.

Fig. 3 : Product lactose values during storage for the period of 42 days. Milk was injected with Group 2 cultures in the quantities of 1 %, 2, 15 % and 3,25 %.

Fig. 4: Graphic representation of dependency of product lactose on the storage period. Samples of Group 1 cultures in the concentration of 1 %, 2, 15 % and 3,25 % are designated as follows: 1 1 %, 1 2% and 1 3 %. Samples of Group 2 cultures in the concentration of 1 %, 2,15 % and 3,25 % are designated as follows: 2_1 %, 2_2% a 2_3 %.

Fig. 5: Product lactose values during storage period of 42 days. Milk was injected with composite culture in the quantities of 1 %, 2, 15 % and 3,25 %.

Fig. 6: Graphic representation of dependency of product lactose on the storage period. The following cultivation environment was monitored: Composite culture injected in milk in the concentration of 1 %, 2,15 % and 3,25 %. The results indicated that the content of lactose ranges around 1,3 % of weight and that increase of input concentration of cultivating cultures has no effect on residual content of lactose in the product. With storage at 4°C, content of lactose may be declared by the age of the product.

Fig. 7: Product lactose values during storage period of 7 days. Cow's milk contained 70% of ewe's milk injected with 3% of the composite culture of Group 1 and Group 2.

Fig. 8: Product lactose values during storage period of 7 days. Cow's milk contained 70%) of ewe's milk injected with 3%> of Group 1 cultures.

Fig. 9: Values of viable bacteria during storage at 4°C for the period of 42 days. Fig. 10: Graphic dependency of the content of viable bacteria on the storage period. 2% of the composite culture of Group 1 and Group 2 was used.

Fig. 11 : Product lactose values during storage period of 27 days. Cow's milk contained 50% of ewe's milk injected with 3% of the composite culture of Group 1, Group 2 and Group3; Group 3 constituted 10% of the total content of cultures.

Fig. 12: Graphic representation of dependency of lactose quantity in the product on the storage period. Cow's milk contained 50% of ewe's milk injected with 3% of the composite culture of Group 1, Group 2 and Group 3; Group 3 constituted 10% of the total content of cultures.

Examples of execution of the invention

Example 1

Pasteurised ewe's milk and cow's milk was poured in a sterile manner in 250ml sterile glass bottles with caps, 100ml each. Milk was tempered to 40 °C. Inoculation composite milk culture was prepared in milk consisting of cow's and ewe's milk in the ratio of 3 :7. The composite culture contained:

two parts of Group 2 cultures: Lactobacillus delbrueckii subsp. bulgaricus and Strepthococcus thermophilus CCM 8532 in the ratio of 1 : 1

one part of Group 1 cultures: Lactobacilus acidophilus CCM 8534 and Lactobacilus helveticus CCM 8533 in the ratio of 1 : 1, total quantity being 1,1.10^s CFU/g.

The prepared inoculation composite milk bacterial culture was injected in the bottles with the prepared milk in the quantity of 1 %, 2, 15 % and 3,25 %. The bottles were then stored in a thermostat tempered to 40°C for the period of 12 hours. Subsequently, the bottles with products were relocated to a cooling box and stored at the temperature of 4°C. Product samples were regularly taken for analysis of the content of viable colonies and the content of lactose.

At the beginning of cultivation, milk contained 4,35% of lactose; after 7 days, lactose content dropped to 1.19 %. The data indicate that this composition of cultures is highly favourable with regard to gradual reduction of lactose content in the product and that the factor of milk composition, or the content of ewe's milk, respectively, is of high significance. The cultures remain viable beyond the period of 42 days of storage in a cold environment.

Example 2

Pasteurised ewe's milk and cow's milk was poured in a sterile manner in 250ml sterile glass bottles with caps, 100ml each. Milk was tempered to 40 °C. Inoculation composite milk bacteria culture was prepared in milk consisting of cow's and ewe's milk in the ratio of 1 : 1. The composite culture contained:

The prepared inoculation composite culture was injected in the bottles with the prepared milk in the quantity of 1 %, 2,15 % and 3, 25 %. The bottles were then stored in a thermostat tempered to 40°C for the period of 12 hours. Subsequently, the bottles with products were relocated to a cooling box and stored at the temperature of 4°C. Product samples were regularly taken for analysis of the content of viable colonies, the content of lactose, lipids and proteins for the period of 42 days.

At the beginning of cultivation, milk contained 4,02 % of lactose; after 7 days, lactose content dropped to 1,5 % and continued to decrease slightly to 1,3 % in the following weeks. The data indicate that this composition of cultures is highly favourable with regard to gradual reduction of lactose content in the product. The cultures remain viable beyond the period of 42 days of storage in a cold environment. This combination of cultures achieves the lowest residual values of lactose content in the product.

The product was fed to dogs in the quantity of up to 200g per 6kg of weight of the dog without symptoms of any negative effect on their digestion.

Example 3

Pasteurised ewe's milk and cow's milk was poured in a sterile manner in 250ml sterile glass bottles with caps, 100ml each. Milk was tempered to 40°C. Inoculation Group 1 culture was prepared in milk consisting of cow's and ewe's milk in the ratio of 1 : 1. The Group 1 culture contained:

Lactobacilus acidophilus CCM 8534 and Lactobacilus helveticus CCM 8533 in the ratio of 1 : 1, total quantity being 1,25.10⁸ CFU/g

The prepared inoculation milk bacteria culture was injected in the bottles with the prepared milk in the quantity of 1 %, 2, 15 % and 3,25 %. The bottles were then stored in a thermostat tempered to 40°C for the period of 12 hours. Subsequently, the bottles with the products were relocated to a cooling box and stored at the temperature of 4°C. Product samples were regularly taken for analysis of the content of viable colonies, the content of lactose, lipids and proteins for the period of 42 days.

At the beginning of cultivation, milk contained 4,2 % of lactose; after 7 days, lactose content dropped to 2,8 % and continued to decrease to 1,8 % in the following weeks. The cultures remain viable beyond the period of 42 days of storage in a cold environment.

Example 4

Pasteurised ewe's milk and cow's milk was poured in a sterile manner in 250ml sterile glass bottle with caps, 100ml each. Milk was tempered to 40°C. Inoculation Group 2 culture was prepared in milk consisting of cow's and ewe's milk in the ratio of 1 : 1. The Group 2 culture contained:

Lactobacillus delbrueckii subsp. bulgaricus and Strepthococcus thermophilus CCM 8532 in the ratio of 1 : 1, total quantity being 1,0.10^s CFU/g

The prepared inoculation composite culture was injected in the bottles with the prepared milk in the quantity of 1 %, 2,15 % and 3,25 % of weight. The bottles were then stored in a thermostat tempered to 40°C for the period of 12 hours. Subsequently, the bottles with the products were relocated to a cooling box and stored at the temperature of 4°C. Product samples were regularly taken for analysis of the content of viable colonies, the content of lactose, lipids and proteins for the period of 42 days. At the beginning of cultivation, milk contained 4,2 % of lactose; after 7 days, lactose content dropped to 2,36 % a continued to decrease slightly to 2 % in the following weeks. The cultures remain viable beyond the period of 42 days of storage in a cold environment. Example 5

Pasteurised ewe's milk and cow's milk was poured in a sterile manner in 250ml sterile glass bottles with caps, 100ml each. Milk was tempered to 40 °C. Inoculation composite milk culture was prepared in milk consisting of cow's and ewe's milk in the ratio of 1 : 1. The composite culture contained:

one part of Group 1 cultures: Lactobacilus acidophilus CCM 8534 and Lactobacilus helveticus CCM 8533 in the ratio of 1 : 1, total quantity being 1,1.10^s CFU/g

0,3 part of Group 3 cultures: Lactobacilus animalis in the quantity of 2,6.10⁷ CFU/g.

The prepared inoculation composite culture was injected in the bottle with the prepared milk 3, 25 %. The bottles were then stored in a thermostat tempered to 40°C for the period of 12 hours. Subsequently, the bottles with the products were relocated to a cooling box and stored at the temperature of 4°C. Product samples were regularly taken for analysis of the content of viable colonies, the content of lactose, lipids and proteins for the period of 42 days.

At the beginning of cultivation, milk contained 4,02 % of lactose; after 5 days, lactose content dropped to 1,9 % and continued to decrease slightly to 1,6 % (and to 1,5 % with 70% proportion of ewe's milk) in the following weeks. The cultures remain viable beyond the period of 42 days of storage in a cold environment.

Industrial exploitability

Animal nutrition, production of milk products, food supplement

Claims

PATENT CLAIMS

An acidophilic yoghurt characterised by being made of an fermented milk consisting of cow's milk with at least 20% share of ewe's milk, containing less than 1,5 % of lactose, at least 10⁷ CFU/g of a composite milk bacterial culture after 6 weeks of storage at the temperature of 4°C; the composite milk bacterial culture contains bacteria of the following strains: Lactobacillus delbrueckii subsp. Bulgaricus and Streptococus thermophilus in the mix deposited under no. CCM 8532, Lactobacillus acidophilus deposited under no. CCM 8534, Lactobacilus helveticus deposited under no. CCM 8533.

The acidophilic yoghurt as per Claim 1 characterised by the composite culture containing bacteria of the type Bifidobacterium animalis or Lactobacillus animalis.

The acidophilic yoghurt as per Claim 1 characterised by the proportion of ewe's milk being 50 % at minimum.

The acidophilic yoghurt as per Claim 3 characterised by containing less than 1,3 % of lactose.

A method of a production of acidophilic yoghurt as per Claim 1 or 2 or 3 or 4 characterised by the pasteurised milk consisting of cow's milk with at least 20% proportion of ewe's milk being tempered to the temperature of 35 to 42°C and injected with 1 to 9 % weight of inoculation composite milk bacterial culture of the minimum concentration of 10⁷ CFU/g; the composite milk bacterial culture contains bacteria of the following type: Lactobacillus delbrueckii subsp. Bulgaricus and Streptococus thermophilus in the mix deposited under no. CCM 8532, Lactobacillus acidophilus deposited under no. CCM 8534, Lactobacilus helveticus deposited under no. CCM 8533; subsequently, the milk is cultivated at the temperature of 38 to 42°C for the period of 10 hours at minimum and the resulting acidophilic yoghurt is then stored at the temperature of 6°C at maximum.

6. The method of production of acidophilic yoghurt as per Claim 5 characterised by the composite milk bacterial culture containing Bifidobacterium animalis or Lactobacillus animalis or their mix.

7. The method of production of acidophilic yoghurt as per Claim 5 characterised by the pasteurised milk consisting of cow's milk with at least 50% proportion of ewe's milk.

8. The method of production of acidophilic yoghurt as per Claim 5 characterised by the milk being inoculated with inoculation composite milk bacterial culture in the quantity of 1 to 3 % of weight.

9. The method of production of acidophilic yoghurt as per Claim 5 or 6 or 7 or 8 characterised by milk cultivation at the temperature of 40°C.

10. The method of production of acidophilic yoghurt as per Claim 5 or 6 or 7 characterised by milk cultivation for the period of 12 hours.

11. The method of production of acidophilic yoghurt as per Claim 5 or 6 or 7 or 8 characterised by storage of the acidophilic yoghurt product at the temperature of 4°C.