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Abstract 1increasing consumer awamess of the potential influence of various foodstuffs on om health. frozen yoghurt traditionally manufacture with yoghurt cultures (St. salivarius ssp. thermophilus + Lb. delbrueckii ssp. bulgaricus) have been though that it consumed not only for their nutritional value but also for their prophylactic properties. But microorganisms of. yoghurt cultures are killed after ingestion by the sever acid conditions in the stomach and the bile juice that is released into the duodenum. Therefore, traditional frozen yoghurt is perceived as a nutritional product that is an excellent source of minerals, carbohydrate and predigested protein. In terms of health benefits, improved lactose-tolerance is the only one that is really well-documented. from these facts, it has become evident that yoghurt cultures are not probiotic bacteria and they may be a limiting factor if traditional frozen yoghurt is used for antibiotic therapy and/or any other medical purpose. These findings have shown the need to use another effective species and strains of beneficial therapeutic effect. Moreover, their metabolites may be responsible for increased shelf-life of the product. by inhibiting a wide spectrum of food spoilage organisms. There fore, the attention subsequently turned to probiotic bacteria. For a culture to he considered probiotic and of therapeutic effect, it must be cabable of survive passage through the upper digestive tract, survive bile salts and growth in the intestine, produce beneficial health effects when in the intestine, and maintain viability and activity in the carrier food before consumption. Once in the intestine, only a limited number of beneficial bacteria can reside there, such as 1. acidophilus and Bifidobacteria. Since some intestinal disturbance occur in the small intestine where 1. acidophilus can survive in abundance, whereas other occur in the large intestine where Bifidobacteria prefer to reside. But these two microorganisms disappear with aging. For this reason, and because of their beneficial therapeutic value, this study aimed to increase numbers of these pro biotic bacteria in ------ .---- ------~_.--- --- ---- 185 human intestinal tract” through consumption them incorporated in a new probiotic fermented ice cream. where their therapeutic and probiotic effects are generally related to inhibition of pathogenic species, reducing the risk of colon cancer, increasing the immune respouse, decreasing concentration of cholesterol iu blood plasma, and improving lactose digestibility for lactose-maldigestors. With these facts in view, the strategy of this study is to make probiotic fermented ice cream of therapeutic value au every day habit for people, and the best way to grow the category is to have people in Egypt understand what a healthy probiotic fermented ice cream. Furthermore, aiming to win over anti-yoghurt people by changing the look and texture of normal yoghurt so that they though they were eating ice cream. Therefore, it was though worthwhile to (1) introduce a new probiotic fermented ice cream product to our market WIder the Egyptian conditions; (2) evaluate the feasibility of incorporating L. acidophilus and Bifidobacteria into this new product insufficient number (>106 cfu/ml) to be considered as pro biotic product of antibacterial role, of beneficial therapeutic value, and acts as protective barrier against invasive pathogens; (3) and observe the effect of pH and long-term frozen storage on survival of these two microorganisms in this new product. This study therefore include the four following parts which might be summarized as follows: Part I: Properties of commercial flavoured frozen yoghurt: In this part, twenty five samples of commercial flavoured frozen yoghurt, representing 5 brands on the Cairo and Giza markets, were randomly collected throughout the summer of 1995, 9 of these samples were labeled as vanilla, 4 as strawberry, 4 as chocolate, 4 as coffee, and the remaining (4) were labeled as mango frozen yoghurt. All samples were analyzed for chemical and microbiological properties. Melting resistance also was determined. The results obtained can be summarized as follows: 1- The majority of samples (640/0) had low titratable acidity, ranged from 0.04 to 0.47%, which are illegal according to the titratable ..- ---------. __.._---------’ 186 acidity which represent generally accepted industry standards in USA (min, 0.5%), On the other hand 52% of samples had a particularly legal titratable acidity values (0.39 to 0.63%) according to the proposed federal standards for I.A. (min. 0.3%). 2- The majority of samples (68%) had a pH level of6.4 to 7.88, which lies with the pH levels of 6.4 to 7.1, which corresponding to the characteristic pH level of standard milk ice and ice cream on the USA markets. 3- The fat content ranged from 3.76 to 5.35% with an average of 4.57%, which lies in the expected range (min.’3.25%) for frozen yoghurt, which represent the generally accepted industry standards in USA. 4- The total solids content of samples ranged from 31.32 to 38.7% (averaged 35.74%), and fall within the expected range (min. 300/0) which represent the generally accepted industry standards in USA. 5- The protein content of samples ranged from 3.53 to 4.52% (averaged 3.890/0).The majority of samples (96%) lies in the range 3.53 to 4.40%. 6- Ash content in the frozen yoghurt examined ranged from 0.69 to 1.04% (averaged 0.83%), which falls within the expected ash content of the commercial ice cream and ice milk (0.7-1.20/0). Samples of higher ash and total solid content, indicating the use of basic formulas of high solids not fat. While samples of high ash content with low total solids content, indicating the use of increased amounts of stabilizer/emulsifier mixture (which contains mineral salts), and/or bases and salts used as pH controlling agents. 7- The total carbohydrates ranged from 19.2 to 26.50/0.These values lie in the expected range of total carbohydrate (20-33%) for ice cream, ice milk, sherbets, water ice and frozen yoghurts. Reducing sugars content ranged from 3.86 to 9.15% in the samples. 8- Acetaldehyde concentration ranged from 2 to 33 ppm. It was higher in mango frozen yoghurt (33 ppm), followed by strawberry (22 ppm); chocolate (18 ppm); vanilla, from NO.5 to 9 (l2 ppm) and the remainder vanilla samples and coffee frozen yoghurt (2 ppm). r--- 187 9- Melting resistance. determined as weight 10ss% at 30°C. ranged from 41.77 to 59.90% and 78.02 to 94.88% after I and 112hr. All frozen yoghurt samples have melted after 2 hr. These variations attributed to the different OVeITlUlbetween samples. Naturally flavouring substances such as strawberry and mango are expected to increase the melting resistance (decrease the weight loss%) of strawberry and mango frozen yoghurt examined than vanilla. chocolate and coffee frozen yoghurt. due to the combined effect of the lower pH values and highly pectin content of strawberry and mango fruits, which consequently decrease amount of the incorporated air in frozen yoghurt. But the higher weight 10850/0was found with mango and strawberry frozen yoghurt after 1!;2 hr (averaged 92.22 and 89.80%. respectively). As might be expected from these findings that strawberry and mango used in the commercial flavoured frozen yoghrut examined were not natural flavouring fruits or juices, but may be artificial flavouring substances. 10- In the present part attempts have been made to specify the product in regard to type of strains used. Total St. salivarius ssp. thermophilus averaged 41.84x105. Corresponding counts for Lb. delbrueckii ssp. bulgaricus averaged 46.2xl05. The results also indicated that, the desired ratio of] : 1 between the two organisms existed only in one sample, No. 14 (40/0).Only 28% of samples had a ratio more than 1 : I in favour of St. salivarius ssp. thermopnilus, while 68% of samples were in favour of Lb. de/brueckii ssp. bulgaricus. 11- The colifonns and presumptive E. coli test have been done for monitoring frozen yoghurt hygiene for public health purpose. Colifonns were detected in all frozen yoghurts examined (ranged from II to 30 cfu/ml, averaged 19.1 cfulml) and exceeded the Egyptian coliform standards of less than ]0 cfu/g, with E. coli absent from 1 g. Only 6 samples (24%) contained E. coli. The high counts of colifonns and presence of E. coli in some samples indicate unfavorable conditions during production, heat treatment under taken might be insufficient and/or recontamination occurs 188 after heat treatment, but the final selling points may also be involved. 12- The yeasts and moulds were detected in all frozen yoghurt examined. It ranged from 10 to 63 cfu/ml (averaged 38.76 cfu/ml). These values were found to lie between the accepted range (l0-50 cfu/ml) and the acceptable range, but to be improved upon (50-100 cfu/ml) suggested by Sames et al. (1979). These findings again indicate unfavourable conditions during production, insufficient heat treatment and/or recontamination (caused by fruits, nuts and sucrose). The final selling points may also be involved. from the results of this part, it is difficult to understand how any commercial frozen yoghurt can show pHs as high as 7.88 or titratable acidities as low as 0.040/0, particularly when frozen yoghurt are classified as acid foods. Such a condition theoretically can be created through pH controlling agents or by an alternative practice might be simply that of adding a yoghurt culture and then not incubating the inoculated mixes. Therefore commercial flavoured frozen yoghurt, abnormally high in pH, which possess low lactic acid bacteria, if incubated, may permit contaminating bacteria to grow to large numbers and pose potential health hazard, as for example, all frozen yoghurts tested had high COlli1tsof coliforms and presence of E. coli in some frozen yoghurts (24%). Therefore, there is no value which may be easily attained in the frozen yoghurt without developing enough lactic acid. These results have shown the need to use an other effective species and strains produce metabolites, to produce frozen yoghurt of increased shelf-life of antibiotic role (against many pathogens, even with frozen yoghurt of high pH and possess low lactic acid); of heneficial therapeutic effect (has a favorable influence on the endogenous intestinal flora); and able to act as protective barrier against invasive pathogens. --- - ----_._- 189 Part II·A new methods for the manufaeatureof Probiotic fermented or aeidi(i~d ice cream of therapeutic yalue , The objectives this part to (1) intorduce a new probiotic fermented or acidified ice cream product to our market under the Egyptian conditions ; (2) evaluate the feasibiltiy of incrorpoarting L.acidophilus and Bifidobacteria Bb- 12 into this new product in sufficient number (> 10 6 cfu / ml ). to be considered as prodiotic fermented or acidified ice cream , of antibacterial role.of beneficial therapeutic value .and acts as protective barrier against invasive pathogens’ (3) observe the effect of pH and long frozen storage on survival of L.acidophilus and Bifidobacteria in this new product. Processing methods of frozen yoghurt discribed by laroia and Martien (1991) are time consuming and icnreased direct and indired production costs .Therefore a new technological aspects were introduced in this part to minimizing of time -consuming and decreasing direct and indirect production costs . These new methods are depending on the’ economically substitition of youghurt culture by two options involved into the following two sections : Sectjon A· The manufacture of ProbiQtic acidfied ice cream by the direct acidification methodt Three distinct mixe were acidified by lactic acid to PHs 6, 5.5 and 5 .Another three distinct mixes were acidified to the same PHs unsing glucono- delta-lactone.All mixes were used for the incrorporation of L. acidophilus and Bifidobacteria The results obtained can be summarized as follows: (1) In all treatments ,decreasing mix - pH results in an increase in the specific gravity and weight by volume , but with comparatively higher values with lactic acid treatments .This could be attributed to decreased whipping rate ,consequently derceased amount of incorparated air in lactic acidacidified ice cream . 2- Decreaing mix.. PH resulted in a decrease amount of incorporated air, but with slightly decreased overrun perrentage with lactic acid -acidified ice cream treatments . 3- Total carbelohydratein all treatments lies in the expected rang of total carboydrate (20 - 30 % ) for frozen yoghurt, and which represent the generally accepted industry standards in USA. -_ ..._-------- ._----_._- - ---’ 190 4- Titratable acidities of lactic acid treatments of PHs 6. 5.5 nd 5 were 0.22 %,0.36 % and 0.51 % ,while it were 0.23 % 0.38 % andO.54 % in GDL_acidified treatments, respectively. 5- Althrough ,all treatments contain most milk ingredients required by Liacidophilus and Bifidobcteria for producing acetaldeyde ,they were not exhihited any activity for producing acetaldehyde.This may be due to that .added acids (Lactic cid or GDL) apear to play and important role in the disturbacne of the endogenous specific metabolic activity of these two organisms or they may be need 10...;,ngetirme for adaptation : 6- In all treatment as pH decreased, melting resistance increased (weight loss % at 30°C decreased) but, with comparatively higher weightloss % in GDL acidifed treaments than that obtained with lactic acid-acidified treatments . 7-Counts of L.acidophilus and Bifidobacteria in all treatments were always in the same log cycle (N x 10 7 cfu / ml ) , duringfrozen storage indicating that these two microorganisms survived in sufficient numbers to ensure thier transit through the stomach ,to produce desirable effects ,when these products used as probiotic foods of therapeutic value. In generally ,the results revealed that, lactic acid -or GDL - acidified treatments showed significant interation differences between treatments of PHs 6 and 5 and length of frozen storage period (up to 8 wx) on counts of Lactdophilus or Bifldobacteriajheretore ,the preferable trend was to probiotic acidified ice cream treatments of pH 6 and pH 5.5 . 8- Sensory evaluation indicats that, these products had acceptable flavour, body / textuer, good melting properties and best palatabity especally for probiotic ice cream treatments of PHs 6 and 5.5 acidified with lactic acid. Section B; The manufacture of Prodiotic fermented ice cream by the indirect method The indirect mothed, in which a fermented mix based on incorporating Bifidobacteria and Lacidophilus cultures only and a pasteurised ice mix are produced separatel9 .These two preparations are then mixed and standardized to pH 6 or 5.5 .based on over all acceptance achieved with direct- acidification .The indirect method was carried out , to evaluate the preferable method to used in the manufacture of probiotic fermented ice cream . The result obtained can be summarized as follows : .. - _.- -_ .. _- - - _. ~ .._. _.. -- -- _ .._.. - 191 1- As PH of the product decreased, specific gravity and wieght by volume icreased .due to decreased whipping ability , consequently decreased amount of incorportated air. 2- Decreasing PH results in a decrease in the amount of incorporated air consequently overrum 0/0. 3- Total carbohydrate in all treatments ( 21.24 - 22.03 %) lies in the expected range of total carbohydrates (20-33 0/0) for ice cream products and frozen yoghurt ,which represents the generally accepted industry standards in USA. 4- The titratble acidity was 0.25 % and 0.38 % for probiotic fermented ice cream of PHs 6 and 5.5 respectively. 5- As PH of the product decreased, acetaldhyde . concentration decreased. 6- Decreased PH (PH5.5 ) does decrease the melting rate of probiotic fermented ice cream of PH 5.5 . 7- Counts of L.acidophilus and Bifidiobacteria were always in the same log cycle ( N x 10 7 cful ml ) during frozen storage .but with comparatively higher counts than those obtained in probiotic acidified ice cream made by directacidification (Section A) up to 7 wk of frozen storage. 8- These results indicated that, the two organisms was servived, in sufficient numbers in probiotic fermented ice cream .to ensure their transitthroug the stomach .to produce desirable effects .when these products used as probiotic food of therapeutic value . 9 Sensory evaluation, indicated that .althrough there are slightly differences between the treatments of PHs 6 and 5.5 as regards flavour, melting quality and body / texture .these products had acceptable flavour, body/texture melting properties and best palatability , especially for probiotic fermented ice cream of PH 6. Part ill. Antibacterial activity of probiotic fermented [acidified ice cream. It was though worth while to investigate the inhibitory role of pro biotic acidified or fermented ice cream treatments of PHs 6 and 5.5 against some spoilage and pathogenic bacteria ,there after ,this study was extended to demonstrate the role played by orgnic acids (Lactic, acetic an gluconic acid created by the hydrolysis of GDL). against the same spoilage and pathogenic bacteria used in this study . The results of this part can be summarized as follows: 192 1- Inhibito activi 0 culture cell -free filttrates of probiotic fermented or acidified ice cream treatments a- Indirect method! The inhibitory effect of soft and hard treatments of PHs 6 and 5.5 was more efficient on staph aureus followed by staph epidermides , Bacillus cereus , salmonella typhi and Escherichia coli, but with comparatively greater effect with soft treatments . Many hypothese have been advanced to explaine this inhibitory effect: (1) production of acids ’ (2) production and elaborations of antibacterial bacteriocins ’ (3) production of short chain fatty acids. h-Direct acidification method by lactic acid or GDL Although, L.acidophilus and Bifidiobacteria survived in sufficient numbers ( N x 10 7 ) in pro biotic acidified ice cream of PH 6 orPH 5.5, cell _ free filtrates of these treatment in the soft or hard form had not any potential to inhibit the five test organisms .This phenomenon may be attributed to that ,directacidification by lactic acid or GDL apears to play an important role in the inactivation (disturbance of) the endogenous specific metabolic activity of the two organisms thus they may be need, longer time for ~dabption . 2- InhibitoO’ effect of Lactic acid or GDL of different PH yalue~ The inhibitory effect of lactic acid or GDL solutions at different PH levels ( 6, 5.5 ,5, 4.5 ,4,3.5 and 3 ) was studied- Only lactic acid solution of PH 3 had a bacteriocidal potential against test organisms. The inhibitor ~ action was greater on salmonella typi .followed by ~ epidermtds , staph aureus , Escherichia coli and Bacillus cereus . 3- Inhibitmy effect of acetic + lactic acicJ solution (3.2) at differentfH levels ( 6. 5.5 . 5,4.5 • 4 , 3.5 and 3 ) The fermentation of two moles of glucose by genus Bifidobacteria leads globally to 3 moles acetate and 2 moles lactate .Therefore ,acetic (PKa= 4.75 ) and lactic (PKa= 3.08 ) acids are among the inhibitors produced by Bifidobacteria .Therefore ,this part is extented to study the inhibitory role played by these acids at different PH levels .against the same test organ- Isms. The results revealed that, acetic + lactic acid s solutions of PH 5,4.5 . 4 , 3.5 and 3 have an inhibitory action on all the test organisms but in different degrees ,with the exception ofsoultaion of PH 5 on staph. aureus , Howev- ------ ------- ---_ ..._------ 193 on er the inhibtitory effect of solution of PH 3 was more efficientltest organisms , followed by solutions of PHs 3.5 , 4, 4.5 and 5 from this part it could be concluded the following : Neither Lacidophilus nor Bifidobacteria (Bb- 12) were effective against test organisms ,when incorporated in acidified ice cream treatments of PH 6 or 5.5 .But probiotic fermented ice cream of PH 6 or 5.5 were more effective on test organisms . On the other hand neither lactic acid GDL nor lactic + acedic acid solutions (2 :3) of PH s 6 orS.5 were effective against test . orgaisms . Therefore ,the inhibitory role of probiotic fermended ice cream of PH 6 or5.5 fermented by L.acidophilus and Bifidobacteria is attributed to proc clueing bacteri,*ns and short chain fatty acids. Finally ,it could be recommended that ,the manufacturing procedure mentioned here in before under section B is the preferable technique to make a probiotic fermented ice cream of a therapeutic value . Part IY Nutritive and therapeutic yalue of probiotic fermentedjce cream. Some socities rely heavily on milk to supply certain nutrients to human infants ,children and adults ,thus this part was carried out to study the ability of probiotic fermented ice cream treatments of PHs 6 and 5.5 to supply the nutritional needs of human children and adults compared with human and cow’s milk .The principle approach has been to relate composition of these treatments to nutrient requirements of human children and adults . 1- Energy yalue and nutrients a- Treatment of PH 6 contained 2.36 and 2.17 time a much calories than does cow’s and human’s milk respectively, while treatment of PH 5.5 contained 2.38 and 2.18 times in the same order. b) The contribution made by100 g treatments of PH 6 or 5.5 (11.97 0/0 or12.0/0 for children and 5.35 or 5.38 % for adults) to the total caloricfic dalily requirements is markedly higher than those provided by human or cow’s milk(5.1 % or 5.15 % for children and 1.11% or 2.47 % for adutls . c) The nutritional adepuancy of the these treatments ,as a pofile relating the content of various constituents to requirements , using energy as common denominator was designated” nutrient density” . Thus for examle -,human children requires about 1.23 g protein per 100 calories .Probiotic fermented ice cream of PH 6 with 4.2 g protein and 155.2 194 calories per 100 g product, provides 4.2/1.23 or 3.42 times the amount of protein needed in relation to energy (per 100 calories) Comparison of the profiles of human and cow’s milk with fermented ice cream treatments of PH 6 or 5.5 in meeting the children or adults requiremnts per 100 calories . In this .for human children or adults ,fermented ice cream furnishsed important excesses of protein and minerals over energy , and hence it could be regarded as perfectly blanced food stuff particularly in relation to human and cow’s milk. 2- Minerals; a- Macrominerals. Those needed at levels of 800 mg or more daily.of which calcium, phosphrous are of important in dairy products . l-Calsium,which esential for bone mineralization as well as for other vital physiological proesses, is the most abundant mineral in the body. Fermented ice cream ofPH6 and 5.5 is one of the most dietary soure ofcalcium ( 157 .14 ) and 155.11 mg/100 /g product respectively) not only because of the significant quantity of calcium present but also because of its ca: ph ratio (1.3 : I ) which is conductive to optimal skeletal growth ’ and the presence of nutrients such as lactose, lactic and acetic acids ,whcih promote C absorption. The contribution made by 100 g treatment of PH 6 is 19.64 % of daily requirement of children and adults. While it was 19.39i’for children and adults with treatment of PH 5.5 . 1- Phosphorus , in combination with calcium is important for bone mineralization .as well as for many chemical reactions in the body. Fermented ice cream treatments contribute from 14.92 - 15.11 % phosphorus of the daily requirements of children and adults, per 100 g . B. Electrolytes : Sodium and potassium are electrolytes, for which the Food and Nutrition Board has estimated safe and adequate daily dietary intakes for children (650 mg ofNa and 1100 mg ofpostossiumlday) and adults (2200 mg and 3250 mg / day respectively). 1-The relationship of sodium intake to hypertension or high blood pressure .a primary risk factor for coronary heart disease, has become an issueof increasing concern in the world .Thus , FDA (1984 ) issued a regulation to make information on the sodium content of foods as a mandatory item in _.-------_.---- -_.- -- _._- ---- ._- --- -_. --- ._-,,- -- - ,.--- - ----- ,._-_. -- 195 in nutrition labelling . In this fermented ice cream of PH 6 (100 g) furnishes es 12.54 % for children and 3.70 % for adults of the total daily requirements While treatment of PH 5 .5 (100 g) provides 11.31 % for children an 3.3,4% for adults of their total daily requirements. •. 2- Preliminary evidence suggests that potassium may protect against a sodium -indcued increase in blood pressure. In this fermented ic cream of PH 6 (IOOg) provides 17.14 % for children and 5.80 % for adults of their daily requirements. While treatment of PH 5.5 furnishes 16.92 % for children and 5.73 % for adults of their daily requirement. 3~Protein content: (a) Probiotic fermented ice cream treatments have a high concentration (4.02 ~4.2 % ) of protein than cow and humans milk. The contritbution made by 100 g treatment of PH 6 (26.25 % and6.67 %) or 100 g treatment of PH 5.s (25.13 % and 6.38 0/0) to the total protein daily requirements of children and adults, respectively is comparatively higher than those provided by cows milk , 20 % and 5.1 % or human milk, 5.63 % and 1.43 % ,for children and adults respectively. b~ Nutritional quality of dietary protein depends largely on the pattern and concentration of essential amino acids provided for the synthesis of nitrogen -containing compounds within the body . In this , fermented ice cream of PH 6 or 5.5 contains, in varying and higher quantities than does cow’s milk all of the amino acids required by human, Moreover ,the pattern of distribution of amino acids in these treatments resmbles that needed by human children and adults . c- Treatment of PH 5.5 , furnishes important excesses of all essential amino acids for children ,and to lesser extent for adults . But, treatment of PH 6 provides lesser contribution for children and adults (per 100 g) . All treatments furnish important excesses of all nonessential and essential amino acids than does cow’s milk. d- Treatments of PH 6 and 5.5 furnish important excesses of glutamic acid (3236.2 and 3342. 11 mg / 100 g respectively) and aspartic (612.26 and 1011.56 mg /100 g respectively) than does cow’s milk. Aspartic and glutomic acids are said to promote the secretion of gastric juices. e~All treatments have a surplus of sulphur -essential amino acids ,which make them valubale in supplementing vegetable protein in the diet which is some what deficient in sulphur amino acids , in particular where protein - ------ -- - - - - --- ---- 196 energy malnutrition is found among children whose diet consists largely of cereals. 4- Fat content. The inabitlity of the animals body to synthesise certian higher unsatuacids raded fatty acids was demonstrated . These certian unsaturated fatty{are essential for normal nutration and for tissue metabolism. They are essential precursors of prostaglandins and thromboxanes, and essential because they are a dietary requirement- linoleic, linolenic an arachidonic are the essentiol fatty acids ( EFA) .The requirement of humans for ~FA has been thoroughly documented .1’ he minimum EFA .requirements’ has been estimeted to be about 0.6 % of the total calories of the diet (65 mg I kcl ) . In this EFA in 100 g of probiotic fermented ice cream of PH 6 ( 0.1668 g EFAand 155.2 calories) contains only enough .t~.account 0.97 ::: 1 1 % of . The total energy in 100 g sample ,.~hire:EFA” in 100 g of treatment of PH ’5..5(cont~ins 0.13 g EFA and 156.1 calories },.co~tains lessen EFA to acco,- -unt 0.75 XJOft~etotal energy in ’lOOg sample .’ In general ,probiotic fermented ice cream treatments are protective products aganist sumptoms of EFA -deficiency because they contain EFA enough to account for 0.6 % of the total energy of each. |