FIELD OF THE INVENTION

The present invention relates to the field of food industry. The invention particularly discloses probiotic fortified food products, particularly milk products. The invention discloses probiotic fortified ghee composition and method of preparation thereof. Invention discloses a ghee preparation or composition as a solution for boosting the immunity.

BACKGROUND OF THE INVENTION

Probiotics are live microbes that can be formulated into many different types of product, including foods, drugs, and dietary supplements. Species of lactobacillus and bifidobacterium are most commonly used as probiotics, but the yeast Saccharomyces cerevisiae, some E.coli and bacillus species are also used as probiotics. Lactic acid bacteria including lactobacillus species, which have been used for preservation of food by fermentation for thousands of years, can serve a dual function by acting as agents for food fermentation and, in addition, potentially imparting health benefits.

US20120093973A1 the invention provides milk-like beverage compositions comprising from about 1 to about 10% of one or more caseinates, from about 1 to about 10% whey protein, from about 2 to about 10% of one or more fats, from about 0.01 to about 1% one or more hydrocolloids, and from about 0.1 to about 10% one or more emulsifiers.

CN1164186C the milk with probiotics contains fat, protein and non-fat milk solid and features that it contains also oligosaccharide in 1-10 w t% and probiotics in 1.0xl0E6 to 3.6xl0E8 each ml and has an acidity of 14-25 titres.

KR101427830B1 the invention relates to immunoreactivity improved fermented milk using probiotics lactic acid bacteria and a production method thereof. Disclosed is a production method of immunoreactivity improved fermented milk using probiotics lactic acid bacteria which comprises a step of injecting Lactobacillus sp YH KCCM 10887P and ABT-5 lactobacillus to raw milk at the same time for fermenting. W002/102168A1 A process for producing extended shelf-life ready-to-use milk compositions containing probiotics by ultra pasteurizing a milk composition, cooling the composition to about 20 to 30 DEG C, and inoculating the composition with one or more aseptically prepared probiotic cultures.

RU2608498C2 invention relates to dairy industry and can be used in production of sour cream butter. Method comprises pasteurizing cream, cooling, ageing cream, whipping, adding to obtained layer a bacterial concentrate of pure cultures Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus diacetylactis, Propionibacterium shermanii in ratio 2: 1:1:1 respectively in an amount of 3-5 % and vegetable oil in an amount of 1.5-3 % to weight of oil with subsequent stirring at temperature of 30-32 °C for 5-10 minutes and cooling.

WO201199875A1 Use of Lactobacillus rhamnosus HN001 or derivatives thereof to treat or prevent rhinitis wherein the Lactobacillus rhamnosus HNOOl or derivative thereof is administered separately, simultaneously or sequentially with one or more agents selected from one or more probiotics, one or more prebiotics, one or more sources of dietary fibre, one or more galactooligosaccharides, one or more short chain galactooligosaccharides, one or more long chain galactooligosaccharides, one or more fructooligosaccharides, one or more short chain galactooligosaccharides, one or more long chain galactooligosaccharides, insulin, one or more galactans, one or more fructans, lactulose, or any mixture of any two or more thereof. Wherein the food comprises cultured milk, yoghurt, cheese, milk drink or milk powder.

US20190015464A1 a foodstuff comprising a substantially anhydrous fatty food, wherein the substantially anhydrous fatty food is solid at room temperature and comprises a substantially anhydrous probiotic bacterial preparation dispersed therein; wherein the fatty food is selected from the group consisting of culinary nut butters, culinary seed butters, culinary bean butters, anhydrous animal fats, anhydrous vegetable fats, anhydrous vegetable oils, and mixtures thereof.

WO2020239761A1 invention relates to compositions and methods for producing fermented milk products with increased amount of probiotic bacteria. In particular, the invention relates to a process for producing a fermented milk which comprises adding to a milk base (i) a starter culture comprising at least one lactose-deficient Streptococcus thermophilus strain, which is capable of metabolizing a non-lactose carbohydrate, and at least one lactose -deficient Lactobacillus strain, which is capable of metabolizing a non-lactose carbohydrate, (ii) one or more non-lactose carbohydrate capable of being metabolized by the lactic acid bacteria, in an amount measured so as to become depleted when the pH of the fermented milk product is between 4.9 and 5.5 and (iii) a probiotic strain selected from a Lactobacillus strain and a Bifidobacterium strain. In addition, the present invention relates to compositions and to fermented milk food or feed products produced by the process of the invention.

Zagazig J. Agric. Res., Vol. 46 No. (6A) 2019 article titled “An attempt to produce synbiotic buttermilk beverages supplemented with barley and oat flour” by Mohamed E. Abd El-Hameed, E. Abd El-Sattar, S.A. Khalifa and A.A. El-Neshawy, study revealed that supplementation with barley and oat flours (2 and 4% for both) showed changes in buttermilk acidity and pH. Whey separation was lower and viscosity was higher in buttermilk sample containing 4%barley and oat flours.

College Dissertations and Theses. 369. https://scholarworks.uvm.edu/graddis/369, Article titled “Chemical Composition, Probiotic Survivability and Shelf Life Studies of Symbiotic Buttermilk” by Dong Zhang, The objective of this study was to develop a symbiotic cultured buttermilk, containing inulin as a prebiotic and the probiotics Lactobacillus acidophilus and Bifidobacterium spp. The cultured buttermilk was prepared using a commercial mesophilic starter CHN22 (Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. lactis, Leuconstoc mesenteorides subsp. cremoris, Lactococcus lactis subsp. lactis biovar diacetylactis) and the probiotics.

Milk and Dairy Products in Human Nutrition: Production, Composition and Health, hirst Edition, Article titled “Butter, Ghee, and Cream Products” by Hae-Soo Kwak, Palanivel Ganesan, and Mohammad A1 Mijan, the purpose of this chapter is to focus on the manufacture and nutrition of butter, ghee, and cream with major emphasis on their health beneficial compounds.

OBJECT OF THE INVENTION

It is main object of the present invention to provide a fortified ghee or butter-oil, also known as clarified butter, drawn butter, and Ghee composition.

It is another object of the present invention to provide a process for preparation of ghee composition comprising of probiotics. It is another object of the present invention to provide a ghee composition that has an advantage to help in boosting / improving immunity by acting in multiple ways.

SUMMARY OF THE INVENTION

One or more problems of the conventional prior arts may be overcome by various embodiments of the present invention.

It is the primary aspect of the present invention to provide a fortified ghee composition, comprising of: clarified butter - 10 g; and probiotics - 500 million - 20 billion microorganism/ colony forming unit.

It is another aspect of the present invention to provide a fortified ghee composition, wherein the probiotics are selected from Lactobacillus rhamnosus, Bifidobacterium lactis, Lactobacillus acidophilus, Streptococci, Lactococci, Leuconostoc, Lactobacillus and Bacillus coagulans.

It is another aspect of the present invention to provide a process for preparation of fortified ghee composition, comprising of steps: procuring and dumping of butter in melting vat; melting of butter in a temperature range of 60 - 80±4°celsius; pre-stratification comprising draining of curd and water; obtaining of ghee or clarified butter and boiling at 110 °C; pre-filtering of ghee; heating at a temperature of 106 °C by continuous mixing or agitation; addition of probiotics, at a temperature range of 68-75 °C; and aging of ghee at temperature range between 15 and 16 °C for up to 24 hours, wherein the concentration of probiotics is probiotics at range between 500 million to 20 billion colony forming units per 10 g of clarified Butter.

It is another aspect of the present invention, wherein the Probiotics is selected from Lactobacillus rhamnosus, Bifidobacterium lactis, Lactobacillus acidophilus, and Streptococci, Lactococci, Leuconostoc, Lactobacillus and Bacillus coagulans, wherein the probiotics is added at range between 500 million to 20 billion colony forming units per 10 g of clarified butter.

It is another aspect of the present invention to provide a process for preparation of fortified ghee composition, wherein the clarified butter is prepared from churned pasteurized milk cream with added butter serum.

It is another aspect of the present invention to provide a fortified ghee composition, wherein the clarified butter comprises of milk fat and milk solids.

It is another aspect of the present invention to provide a fortified ghee composition, wherein the milk fat is obtained from fresh cream from milk which is free from any additives, preservatives or adulterants.

It is another aspect of the present invention to provide a fortified ghee composition, wherein the package temperature for fortified ghee is 45 °C.

It is another aspect of the present invention to provide a fortified ghee composition, wherein the storage temperature for fortified ghee is 20±2 °C.

BRIEF DESCRIPTION OF DRAWINGS

So that the manner in which the features, advantages and objects of the invention, as well as others which will become apparent, may be understood in more detail, more particular description of the invention briefly summarized above may be had by reference to the embodiment thereof which is illustrated in the appended drawings, which form a part of this specification. It is to be noted, however, that the drawings illustrate only a preferred embodiment of the invention and is therefore not to be considered limiting of the invention's scope as it may admit to other equally effective embodiments.

Figure 1 illustrates the flow diagram of method of preparation of probiotic based clarified butter or ghee according to one embodiment of the present invention.

Figure 2 illustrates process flow chart for cow and buffalo ghee according to an embodiment of the present invention.

Figure 3 illustrates graph of % survival of the probiotics organism at the end of 36 months. Figure 4: illustrates graph of % survival of the probiotics organism in gastric juice at the end of 2 hours or 120 min.

Figure 5 : illustrates graph of % survival of the probiotics organism in intestinal juice at the end of 2 hours or 120 min.

DETAILED DESCRIPTION OF THE INVENTION

It is to be understood that the present disclosure is not limited in its application to the details of construction and the arrangement of components set forth in the following description or illustrated in the drawings. The present disclosure is capable of other embodiments and of being practiced or of being carried out in various ways. Also, it is to be understood that the phraseology and terminology used herein is for the purpose of description and should not be regarded as limiting.

The present invention relates to fortified ghee i.e. ghee containing probiotics. Major advantage of this composition is that it helps in boosting / improving immunity by acting in multiple ways.

The following specification provides relevant technical guidelines of probiotic ghee of the present invention.

I. Cow ghee

II. Buffalo ghee

General guidelines on manufacturing activities in dairy plant

Milk reception is critical as this is the first step from where the further process starts. On arrival of milk tanker, staff verified the voucher. The seal condition on the tanker valves is checked before drawing of milk sample. Visual observation of the milk surface is made for presence of any foreign matter. The surface should be free from foreign matters. The milk sample from tanker valves is observed and should be free from foreign matters.

Table 1: Specifications for mix milk

Table 2: Specifications for cow milk

*Normal temperature of milk supplied shall not exceed 7 °C but occasionally if it goes up to 10 °C, the same will be accepted provided that the quality otherwise is acceptable as per above standards.

** To be carried out periodically. Note: Concentrate milk to be diluted to 8.5 % SNF for analysis.

The incoming milk is accepted only after confirming sample free from presence of adulterants and preservatives. If the quality of milk is not meeting the acceptability norms of FSSAI standards the milk should not be used for the manufacturing of clarified butter or Ghee for FSSAI.

According to one embodiment of the present invention the fortified healthy ghee comprises both probiotics, and beta glucan. The present invention gives low cholesterol ghee, good for immunity, digestion and other health benefits. Ghee with Probiotic Effects i.e. bacterial strains like in a concentration of 500 million to 20 billion microorganisms per 10 g of ghee.

Probiotic strains used: the most common families of bacteria being Streptococci,

Lactococci,

Leuconostoc,

Lactobacillus,

Lactobacillus rhamnosus,

Bifidobacterium lactis, and Lactobacillus acidophilus.

It is clinically proven.

Bacillus coagulans:

B. coagulans is a rod-shaped, slightly acidophilic, gram-positive, catalase-positive, spore forming, thermotolerant, aerobic to microaerophilic, highly resilient probiotic bacteria. Bacillus coagulans strain has been deposited with the Microbial Type Culture Collection (MTCC) -assigned number MTCC 5724 and with Belgian Coordinated Collections of Microorganism (BCCM™ILGM) with the assigned number LMG S -27484.

Stability

A probiotic to be effective it should stay stable until it reaches the target site in the gastrointestinal tract.

Following are the challenges a live probiotic organism undergoes until this process, · Storage and handling until processing

• Processing conditions such as heat, pressure etc.

• Food composition and conditions such as pH and presence of other antimicrobial constituents

• Shelf life and handling until consumption.

• Acidic conditions in the stomach, pH can be below 2

• Bile acids which tend to impact probiotic viability Bacillus coagulans have the key to these challenges. Bacillus coagulans is an aerobic, spore forming, lactic acid probiotic bacteria which has superior stability as compared to the traditional vegetative organisms. This unique quality makes it the best probiotic option from the stability perspective. Safety:

Bacillus coagulans is one of the oldest and most widely used strains. Over 130 million doses prescribed and consumed annually. Bacillus coagulans is considered safe for chronic human consumption and has received United States FDA Generally Recognized as Safe (GRAS) status including Infant GRAS. Bacillus coagulans is of Non-GMO origin and is produced in a GMP, FMIQS and NSF certified facility.

Efficacy:

Bacillus coagulans is supported by twenty plus scientific publications covering gut health, Immunity against GI track pathogens, inhibition of food borne pathogens, inhibition of bacteria causing dental carries & gingivitis and bacterial vaginosis (BV). Gut health, Immunity against GI track pathogens & Inhibition of food borne pathogens Studies cover neonates, pediatrics and adult population in India. Most of them are independently published, peer reviewed placebo controlled. Condition studied include diarrhea, rotaviral diarrhea, gastroenteritis, gas and bloating. Food borne pathogens studied for zone of inhibition include Escherichia coli, Pseudomonas, B. cereus, Salmonella, Staphylococcus aureus. A prospective double -blind randomized study conducted by GI Chandra has shown that feeding Bacillus coagulans on a prophylactic basis in the first year of life has a significant preventive impact on the incidence and duration of diarrhea as well as the total number of days of illness. This 12-month long study concludes a reduction in the number of illness days among treatment group down to 13 from 35 days in the placebo group reiterating immunity or natural defense against GI pathogens such as rotavirus.

Studies conducted by Dhongade, R.K et al, Deodhar, J et al, Dr Balabhai Nanavati and Mukesh Choudhary et al also concludes the role of bacillus coagulans in the GI health.

Inhibition of oral pathogens causing dental carries & gingivitis Oral studies cover Indian children of age group 7 to 14 and adults of 20 - 30 years. These studies include the inhibitory effect of bacillus coagulans on the cariogenic bacteria streptococcus mutans and parameters of gingivitis. A study by A Jindal et al concludes the administration of bacillus coagulans in a group of Indian children resulted in a significant reduction of a cariogenic microorganism, mutans streptococci. With an increasing global problem of antibiotic resistance, contributing to preventing the effective treatment of microbial diseases, probiotic by virtue of their natural therapy appear to be a novel approach for carries prevention. Dietary supplementation of probiotics might be used as a suitable, easy safe method for carries prevention. Study by Pumima Vidyesh Nadkerny et al concludes that probiotics offer a natural and promising option to establish both a good oral and systemic health. The probiotic mouth rinse tested with bacillus coagulans was effective in reducing plaque accumulation and gingival inflammation· Oral probiotics represents a breakthrough approach to maintain oral health by utilizing natural beneficial bacteria to provide a natural defense against those bacteria thought to be harmful to teeth and gums.

Can Reception

On arrival of route vehicles, the cans are unloaded on can conveyor; lids are opened using lid opener or wooden/rubber hammer. Checked for sediment and free-floating fat. Segregated by smell and taste and in case of any abnormal taste or smell such cans are separated or segregated. The milk of segregated cans is subjected to further examination. Milk having abnormal taste, smell, sediment and adulterants if any is discarded.

Milk from all the VLCs is also checked for neutralizer, additives, adulterants and MBRT on rotational basis so as to cover all the VLCs within a specific period. Abnormal milk is rejected and further detail checking is carried out at the society level. The accepted milk is weighed, strained and pumped to storage tank after passing through filter and chiller. The liquid milk is stored at 5°C or below for further processing.

Pasteurization of milk

Milk processing is started through pasteurizer after thorough sanitization at 80°C for 20 minutes. Raw milk is taken in balance tank. During processing, milk passes through 60 -mesh size nylon filter, clarifier and HTST pasteurizer. Pasteurization temperature is maintained at 75±2°C for 15 seconds holding to get better shelf life of milk processed (i.e. MBRT >6 hours). All instrumentation and controls of pasteurizer plant are kept functional. Phosphate test and MBRT of milk, sampled from the plant is carried out in the beginning of every shift during processing to confirm the plant capability for milk processing. Thermograph of previous day is forwarded for I/C QC for his inferences on quality of milk processing.

Ghee production process

The process starts with storage of pasteurized cream at 10°C, addition of butter serum [203], churning for 40-45 min [202] to yield butter. The butter or pumped to the butter-melting vat for ghee manufacture. (Reference Figure 2: A for process flow diagram).

The butter is melted by indirect heating method with the help of steam in coils in the melting vat jackets [205]. The molten butter [205a] is then pumped into the pre stratification tank [205b] at a temperature of 80+/- °C, preferably at a temperature of 70°C, the butter get separated into three layers (Top layer of floating denatured particles of curd, a middle layer of fat and a bottom layer of buttermilk). The lower serum portion is taken out. The rest of butter is pumped to the ghee boiler [206] where it is heated to a temperature of about 107 - 109°C and boiled at that temperature for 15- 20 minutes with agitation followed by pre-filtering [206a] . The ghee is then pumped to the settling tank [207] and sent to the ghee clarifier [207a] through a pair of duplex filters. The clarified ghee is stored in ghee storage tank [210] at a temperature of about 20°C for granulation. Ghee is examined by QC or Quality Control and if fails, sent to the settling tank [207]. Ghee which is approved by QC is sent to ghee filling tank [211], followed by packaging [212] at 45 °C and stored at about 20°C +/- 2°C for granulation. This ghee is sent for packaging [212] and stored [215] at a controlled temperature for grain formation. In this process through between preparation of ghee in boiler [206] and Ghee clarification [206], intermediate steps of separation of coarse berry at pre-filtering stage [206a], Fine berry from settling tank [207] and Fine berry from ghee clarifier stage [207a] is sent for ghee extraction to ghee boiler [206] .

In one embodiment [Figure 2] the ghee from the ghee settling tank [207] is sent chilled [208] and sent [209] for reprocess. The package temperature for fortified ghee is 45°C. The storage temperature for fortified ghee is 20±2 °C. It is finally dispatched at ambient temperature in covered vans [216]. Packaging: ghee from the Ghee filling tank [211], is sent to consumer bulk packaging [212] and left for aging [213] followed by Cartoning and batch coding [214], Sent to finished good store [215] and dispatched [216].

The remaining ghee residue is collected in cans and transferred to a fat recovery vat at the end of the shift. Here the ghee residue is heated with water and then chilled with chilled water [208]. The fat present gets liquefied first and float on top and then it gets solidified. This is separated out. The extracted ghee residue is disposed off.

SOP for cleaning and sterilization

Cleaning and sterilization is a very important aspect of a dairy plant. Cleaning is done mostly in place or CIP, automatically but manual cleaning is also done in certain intervals. The CIP system is multi use type. Mainly used cleaning medium is caustic soda and nitric acid. Their strength is measured by conductivity meter and adjusted automatically. Sanitization is done mainly by hot water but formalin is also used for fumigation. Cleaning system in different section:

Pre - process section:

A. Pasteurizers are cleaned locally by short and long CIP

I. Short CIP:

• Pre rinse with DM water for 15 minutes till flush water is free from milk residue.

• 1% lye solution at 80° C is circulated for 45min.

• Flush lye solution with DM water at 80° C till pH of flush water same as that of DM water.

II. Long CIP:

• Flush with DM water / soft water for 15min and contains till fresh water comes.

• 1% lye solution at 80° C is re-circulated for 30min.

• Flush lye with DM water for 15min

• 1% acid with DM water at 80° C for 15 min.

• DM water and then Lye to pass through again throughout the system

• Flushing lye with DM water till phenolphthalein does not give pink colour

• Hot water at 90° C is circulated for 15-20 min for sterilization.

B. Pipelines, Silos tanks

• Flush with water

• 1% lye solution at 80°C is circulated for 15 -20 min

• Flush lye with DM water.

• Hot water at 90° C / 15min to sterilize

Sometimes they are chlorinated at QAC (200ppm) till next use. Phosphate is used as a chelating agent along with lye to the extent of 0.1 % Hexa-meta phosphate to improve cleaning efficiency and rinsability of lye solution.

Butter and ghee section:

• All pipelines and vat or tanks are rinsed with hot water at 60 - 65°C, 1% caustic solution at 80°C and final rinse to remove alkali and hot water sterilization at 90 -95°C.

Plant maintenance

• Proper preventive maintenance schedule for upkeep of machine and equipment are exercised so as to get trouble free milk processing and packing.

• 2.9 Manpower Development • Trained manpower is kept for milk reception, processing, packing and dispatch as per job requirement.

Advantages and benefits: This ghee while keeping the superior taste and aroma intact, helps in building body immunity and aids digestion. The fortified ghee while keeping the superior taste and aroma intact, helps in building body immunity, aids digestion, lowers cholesterol giving an all-round body boost to fight off diseases. Table 3: The product specification of the present invention (fortified ghee):

In reference to Figures 3, the product is evaluated for probiotics (% survival vs time) the probiotic is active for a period of 36 months or 3 years.

Figure 4, the product is evaluated for % survival in gastric juice 120 min. Figure 5, the product is evaluated for % survival in intestinal juice 120 min.