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l-149\slmult0 \par\pard\ql\li4516\sb81\sl-149\slmult0 \up0 \expndtw0\charscalex1

04 \ul0\nosupersub\cf7\f8\fs13 Food Research International 42 (2009) 1233-1239 \
par\pard\ql \li4570\sb0\sl-184\slmult0 \par\pard\ql\li4570\sb0\sl-184\slmult0 \p
ar\pard\ql\li4570\sb15\sl-184\slmult0 \up0 \expndtw0\charscalex104 \ul0\nosupers
ub\cf16\f17\fs16 Contents lists available at\ul0\nosupersub\cf17\f18\fs16 {\fie
ld{\*\fldinst {HYPERLINK http://www.sciencedirect.com/science/journal/09639969 }
}{\fldrslt {\ul0\nosupersub\cf17\f18\fs16 ScienceDirect}}} \par\pard\ql \li4194\
sb262\sl-322\slmult0 \up0 \expndtw0\charscalex102 \ul0\nosupersub\cf18\f19\fs28
Food Research International \par\pard\ql \li3582\sb0\sl-184\slmult0 \par\pard\ql
\li3582\sb148\sl-184\slmult0 \up0 \expndtw0\charscalex123 \ul0\nosupersub\cf11\f
12\fs16 journal homepage : {\field{\*\fldinst {HYPERLINK http://www.elsevier.com
/locate/foodres }}{\fldrslt {\ul0\nosupersub\cf11\f12\fs16 www .elsevier .com /l
ocate /foodres}}} \par\pard\ql \li850\sb0\sl-218\slmult0 \par\pard\ql\li850\sb21
0\sl-218\slmult0 \up0 \expndtw-1\charscalex100 \ul0\nosupersub\cf1\f2\fs19 Revie
w \par\pard\ql \li850\sb127\sl-310\slmult0 \up0 \expndtw0\charscalex103 \ul0\nos
upersub\cf2\f3\fs27 Ice-cream as a probiotic food carrier \par\pard\ql \li850\sb
197\sl-230\slmult0 \up0 \expndtw-3\charscalex100 \ul0\nosupersub\cf3\f4\fs21 Adr
iano G. Cruz\ul0\super\cf4\f5\fs22 a,\ul0\super\cf5\f6\fs22 *\ul0\nosupersub\cf
3\f4\fs21 , Adriane E.C. Antunes\ul0\super\cf4\f5\fs22 b\ul0\nosupersub\cf3\f4\
fs21 , Ana Lcia O.P. Sousa\ul0\super\cf4\f5\fs22 c\ul0\nosupersub\cf3\f4\fs21 ,
Jos A.F. Faria\ul0\super\cf4\f5\fs22 a\ul0\nosupersub\cf3\f4\fs21 , Susana M.I.
Saad\ul0\super\cf4\f5\fs22 c \par\pard\qj \li850\ri420\sb112\sl-180\slmult0 \up
0 \expndtw-1\charscalex100 \ul0\super\cf6\f7\fs13 a\ul0\nosupersub\cf7\f8\fs13
Departamento de Tecnologia de Alimentos, Faculdade de Engenharia de Alimentos, U
niversidade Estadual de Campinas, Caixa Postal 6121, 13083-862 Campinas, So Paulo
, Brazil \up0 \expndtw-1\charscalex100 \ul0\super\cf6\f7\fs13 b\ul0\nosupersub\c
f7\f8\fs13 Faculdade de Cincias Aplicadas, Universidade Estadual de Campinas, R.
Pedro Zacarias, 1300, 13484-350 Limeira, SP, Brazil \par\pard\ql \li850\sb26\sl
-149\slmult0 \up0 \expndtw-3\charscalex100 \ul0\super\cf6\f7\fs13 c\ul0\nosupers
ub\cf7\f8\fs13 Departamento de Tecnologia Bioqumico-Farmacutica, Faculdade de Cinc
ias Farmacuticas, Universidade de So Paulo, Av. Prof. Lineu Prestes, 580, 05508-00
0 So Paulo, SP, Brazil\par\pard\sect\sectd\sbknone\cols2\colno1\colw3981\colsr160
\colno2\colw7619\colsr160\ql \li850\sb0\sl-207\slmult0 \par\pard\ql \li850\sb207
\sl-207\slmult0 \up0 \expndtw0\charscalex135 \ul0\nosupersub\cf8\f9\fs18 a r t i
c l e i n f o\par\pard\ql \li850\sb0\sl-149\slmult0 \par\pard\ql \li850\sb70\
sl-149\slmult0 \up0 \expndtw0\charscalex102 \ul0\nosupersub\cf7\f8\fs13 Article
history:\par\pard\ql \li850\sb24\sl-149\slmult0 \up0 \expndtw0\charscalex100 Rec
eived 1 January 2009\par\pard\ql \li850\sb23\sl-149\slmult0 \up0 \expndtw0\chars
calex103 Accepted 30 March 2009\par\pard\ql \li850\sb0\sl-149\slmult0 \par\pard\
ql \li850\sb0\sl-149\slmult0 \par\pard\ql \li850\sb126\sl-149\slmult0 \up0 \expn
dtw-1\charscalex100 Keywords:\par\pard\ql \li850\sb24\sl-149\slmult0 \up0 \expnd
tw0\charscalex100 Ice-creams\par\pard\ql \li850\sb24\sl-149\slmult0 \up0 \expndt
w-2\charscalex100 Processing\par\pard\ql \li850\sb23\sl-149\slmult0 \up0 \expndt
w0\charscalex103 Probiotics\par\pard\ql \li850\sb23\sl-149\slmult0 \up0 \expndtw
-2\charscalex100 Lactobacillus\par\pard\ql \li850\sb24\sl-149\slmult0 \up0 \expn
dtw0\charscalex100 Bifidobacterium\par\pard\ql \li850\sb0\sl-172\slmult0 \par\pa
rd\ql \li850\sb0\sl-172\slmult0 \par\pard\ql \li850\sb0\sl-172\slmult0 \par\pard
\ql \li850\sb134\sl-172\slmult0 \up0 \expndtw0\charscalex106 \ul0\nosupersub\cf9
\f10\fs15 Contents\par\pard\ql \li970\sb0\sl-172\slmult0 \par\pard\ql \li970\sb4
0\sl-172\slmult0\tx1305 \up0 \expndtw-2\charscalex100 1.\tab \up0 \expndtw0\char
scalex105 Introduction . . .\par\pard\ql \li970\sb20\sl-172\slmult0\tx1305 \up0
\expndtw-2\charscalex100 2.\tab \up0 \expndtw0\charscalex102 Probiotic ice-cream
processing . . .\par\pard\column \ql \li4141\sb0\sl-207\slmult0 \par\pard\ql \
li4141\sb0\sl-207\slmult0 \par\pard\ql \li20\sb5\sl-207\slmult0 \up0 \expndtw0\c
harscalex129 \ul0\nosupersub\cf8\f9\fs18 a b s t r a c t\par\pard\qj \li4141\sb0
\sl-191\slmult0 \par\pard\qj \li20\ri450\sb10\sl-191\slmult0 \up0 \expndtw0\char
scalex103 \ul0\nosupersub\cf9\f10\fs15 Ice-creams are food products showing pote
ntial for use as probiotic vehicles, with the added advantage of \up0 \expndtw0\
charscalex104 being appreciated by people belonging to all age groups and social
levels. However, the development of \up0 \expndtw0\charscalex105 ice-creams co
ntaining probiotic bacteria requires the overcoming of certain technolog

ical intrinsic \up0 \expndtw0\charscalex105 requirements related to their proce
ssing stages. The aim of the present paper was to review the techno-\line \up0 \
expndtw0\charscalex106 logical parameters involved in the production of probioti
c ice-creams. Although the application of probi-\line \up0 \expndtw0\charscalex1
04 otics in cheeses, and especially in fermented milks, has been widely explored
in the literature, ice-cream \up0 \expndtw0\charscalex106 is a relatively innov
ative matrix for the application of probiotics, and thus a review about its pote
ntial as \up0 \expndtw0\charscalex107 probiotic food carrier could be very helpf
ul.\par\pard\ql \li4567\sb17\sl-172\slmult0 \up0 \expndtw0\charscalex103 2009 El
sevier Ltd. All rights reserved.\par\pard\ql \li970\sb0\sl-172\slmult0 \par\pard
\ql \li970\sb0\sl-172\slmult0 \par\pard\ql \li970\sb0\sl-172\slmult0 \par\pard\q
l \li970\sb0\sl-172\slmult0 \par\pard\ql \li970\sb0\sl-172\slmult0 \par\pard\ql
\li970\sb0\sl-172\slmult0 \par\pard\ql \li970\sb0\sl-172\slmult0 \par\pard\ql \l
i6786\sb46\sl-172\slmult0 \up0 \expndtw0\charscalex103 1233\par\pard\ql \li6786\
sb20\sl-172\slmult0 \up0 \expndtw0\charscalex103 1234 \par\pard\sect\sectd\sbkno
ne \li970\sb20\sl-172\slmult0\fi0\tx1310\tx10907 \up0 \expndtw0\charscalex104 3.
\tab \up0 \expndtw0\charscalex105 Technological hurdles for the incorporation of
probiotic bacteria into ice-cream . . .\tab \up0 \expndtw0\charscalex104 1234\p
ar\pard\sect\sectd\sbknone \li970\sb20\sl-172\slmult0\fi350\tx1782\tx10907 \up0
\expndtw0\charscalex104 3.1.\tab \up0 \expndtw0\charscalex104 Fruit pulp or juic
e as an ingredient of probiotic ice-creams . . .\tab \up0 \expndtw0\charscalex1
04 1235\par\pard\sect\sectd\sbknone \li970\sb21\sl-172\slmult0\fi350\tx1781\tx10
907 \up0 \expndtw0\charscalex104 3.2.\tab \up0 \expndtw0\charscalex104 Addition
of probiotic cultures to ice-cream . . .\tab \up0 \expndtw0\charscalex104 1236\p
ar\pard\sect\sectd\sbknone \li970\sb20\sl-172\slmult0\fi350\tx1781\tx10907 \up0
\expndtw0\charscalex104 3.3.\tab \up0 \expndtw0\charscalex104 Stability of the p
robiotic cultures during ice-cream storage . . .\tab \up0 \expndtw0\charscalex1
04 1236\par\pard\sect\sectd\sbknone \li970\sb20\sl-172\slmult0\fi350\tx1781\tx10
907 \up0 \expndtw0\charscalex104 3.4.\tab \up0 \expndtw0\charscalex104 Overrun a
nd probiotic microorganisms . . .\tab \up0 \expndtw0\charscalex104 1236\par\pard
\sect\sectd\sbknone \li970\sb21\sl-172\slmult0\fi350\tx1781\tx10907 \up0 \expndt
w0\charscalex104 3.5.\tab \up0 \expndtw0\charscalex105 Influence of storage cond
itions on the characteristics of probiotic ice-creams . . .\tab \up0 \expndtw0\c
harscalex104 1237\par\pard\sect\sectd\sbknone \li970\sb21\sl-172\slmult0\fi0\tx1
310\tx10907 \up0 \expndtw0\charscalex104 4.\tab \up0 \expndtw0\charscalex104 Sen
sory features of probiotic ice-creams . . .\tab \up0 \expndtw0\charscalex104 123
7\par\pard\sect\sectd\sbknone \li970\sb19\sl-172\slmult0\fi0\tx1310\tx10907 \up0
\expndtw0\charscalex104 5.\tab \up0 \expndtw0\charscalex104 Perspectives . . .\
tab \up0 \expndtw0\charscalex104 1238\par\pard\sect\sectd\sbknone \li970\sb21\sl
-172\slmult0\fi358\tx10907 \up0 \expndtw0\charscalex104 Acknowledgements . . .\t
ab \up0 \expndtw0\charscalex104 1238\par\pard\sect\sectd\sbknone \li970\sb20\sl172\slmult0\fi358\tx10907 \up0 \expndtw0\charscalex104 References . . .\tab \up0
\expndtw0\charscalex104 1238\par\pard\sect\sectd\sbknone\cols2\colno1\colw6070\
colsr160\colno2\colw5530\colsr160\ql \li850\sb0\sl-184\slmult0 \par\pard\ql \li8
50\sb0\sl-184\slmult0 \par\pard\ql \li850\sb0\sl-184\slmult0 \par\pard\ql \li850
\sb0\sl-184\slmult0 \par\pard\ql \li850\sb179\sl-184\slmult0 \up0 \expndtw0\char
scalex114 \ul0\nosupersub\cf10\f11\fs16 1. Introduction\par\pard\qj \li850\sb0\s
l-209\slmult0 \par\pard\qj \li850\ri149\sb5\sl-209\slmult0\fi232 \up0 \expndtw0\
charscalex104 The development of new food products turns out to be increas-\line
\up0 \expndtw0\charscalex105 ingly challenging, as it has to fulfill the consum
er\ul0\nosupersub\cf11\f12\fs16 \u8217?\ul0\nosupersub\cf10\f11\fs16 s expectanc
y for \line \up0 \expndtw0\charscalex103 products that are simultaneously relish
and healthy. Probiotics are \line \up0 \expndtw0\charscalex108 defined as live
microorganisms that, when administered in ade-\line \up0 \expndtw0\charscalex103
quate amounts, confer health benefits on the host (\ul0\nosupersub\cf12\f13\fs1
6 Food and Agri-\line \up0 \expndtw0\charscalex103 culture Organization of Unite
d Nations; World Health Organization \line \up0 \expndtw0\charscalex102 - FAO/WH
O, 2001\ul0\nosupersub\cf10\f11\fs16 ). Probiotic food is defined as a food prod
uct that \line \up0 \expndtw0\charscalex105 contains viable probiotic microorgan
isms in sufficient populations\par\pard\ql \li850\sb0\sl-149\slmult0 \par\pard\q

l \li850\sb0\sl-149\slmult0 \par\pard\ql \li951\sb18\sl-149\slmult0 \up0 \expndt
w0\charscalex102 \ul0\nosupersub\cf14\f15\fs13 *\ul0\nosupersub\cf7\f8\fs13 Co
rresponding author. Tel.: +55 19 35214016.\par\pard\ql \li1088\sb24\sl-149\slmul
t0 \up0 \expndtw-1\charscalex100 E-mail address:\ul0\nosupersub\cf15\f16\fs13 {
\field{\*\fldinst {HYPERLINK mailto:food@globo.com }}{\fldrslt {\ul0\nosupersub\
cf15\f16\fs13 food@globo.com}}}\ul0\nosupersub\cf7\f8\fs13 (A.G. Cruz).\par\par
d\ql \li850\sb0\sl-149\slmult0 \par\pard\ql \li850\sb62\sl-149\slmult0\tx2937 \u
p0 \expndtw0\charscalex106 0963-9969/$ - see front matter\tab \up0 \expndtw0\cha
rscalex103 2009 Elsevier Ltd. All rights reserved.\par\pard\ql \li850\sb23\sl-14
9\slmult0 \up0 \expndtw0\charscalex102 doi:10.1016/j.foodres.2009.03.020\par\par
d\column \ql \li6230\sb0\sl-184\slmult0 \par\pard\ql \li6230\sb0\sl-184\slmult0
\par\pard\ql \li6230\sb0\sl-184\slmult0 \par\pard\ql \li6230\sb0\sl-184\slmult0
\par\pard\ql \li20\sb179\sl-184\slmult0 \up0 \expndtw0\charscalex104 \ul0\nosupe
rsub\cf10\f11\fs16 incorporated in a suitable matrix (\ul0\nosupersub\cf12\
f13\fs16 Gibson & Roberfroid, 1995;\par\pard\qj \li20\ri450\sb5\sl-209\slmult
0 \up0 \expndtw0\charscalex103 Saxelin, Korpela, & Mayara-Makinen, 2003\ul0\nosu
persub\cf10\f11\fs16 ). This means that their \up0 \expndtw0\charscalex105 viabi
lity and metabolic activity must be maintained in all the steps \up0 \expndtw0\c
harscalex105 of the food processing operation, from their production up to their
\up0 \expndtw0\charscalex103 ingestion by the consumer, and also that they must
be able to sur-\line \up0 \expndtw0\charscalex105 vive in the gastrointestinal
tract (\ul0\nosupersub\cf12\f13\fs16 Sanz, 2007\ul0\nosupersub\cf10\f11\fs16 ).\
par\pard\qj \li20\ri451\sb1\sl-209\slmult0\fi232 \up0 \expndtw0\charscalex105 Th
e information available concerning the concentration of pro-\line \up0 \expndtw0
\charscalex109 biotic microorganisms needed for biological effects leads to the
\up0 \expndtw0\charscalex115 conclusion that it will vary as a function of the s
train and the \up0 \expndtw0\charscalex103 health effect desired (\ul0\nosupersu
b\cf12\f13\fs16 Champagne, Gardner, & Roy, 2005\ul0\nosupersub\cf10\f11\fs16 ).
Never-\line \up0 \expndtw0\charscalex103 theless, populations of \up0 \expndtw
0\charscalex104 10\ul0\super\cf13\f14\fs16 6\ul0\nosupersub\cf10\f11\fs16 -10\ul
0\super\cf13\f14\fs16 7\ul0\nosupersub\cf10\f11\fs16 CFU/g in the final pro
duct are\par\pard\qj \li20\ri451\sb1\sl-209\slmult0 \up0 \expndtw0\charscalex10
7 established as therapeutic quantities of probiotic cultures in pro-\line \up0
\expndtw0\charscalex105 cessed foods (\ul0\nosupersub\cf12\f13\fs16 Talwalkar,
Miller, Kailasapathy, & Nguyen, 2004\ul0\nosupersub\cf10\f11\fs16 ), \par\pard\
sect\sectd\fs24\paperw11900\paperh15860\pard\sb0\sl-240{\bkmkstart Pg2}{\bkmkend
Pg2}\par\pard\li654\sb0\sl-149\slmult0\par\pard\li654\sb0\sl-149\slmult0\par\pa
rd\li654\sb0\sl-149\slmult0\par\pard\li654\sb0\sl-149\slmult0\par\pard\li654\sb1
04\sl-149\slmult0\fi0\tx3916 \up0 \expndtw-1\charscalex100 \ul0\nosupersub\cf7\f
8\fs13 1234\tab \up0 \expndtw-1\charscalex100 A.G. Cruz et al. / Food Research I
nternational 42 (2009) 1233-1239\par\pard\sect\sectd\sbknone\cols2\colno1\colw58
74\colsr160\colno2\colw5726\colsr160\ql \li654\sb0\sl-184\slmult0 \par\pard\ql \
li654\sb58\sl-184\slmult0 \up0 \expndtw0\charscalex102 \ul0\nosupersub\cf10\f11\
fs16 reaching 10\ul0\super\cf13\f14\fs16 8\ul0\nosupersub\cf10\f11\fs16 -10\ul0\
super\cf13\f14\fs16 9\ul0\nosupersub\cf10\f11\fs16 CFU, provided by a daily con
sumption of 100 g or\par\pard\ql \li654\ri149\sb4\sl-209\slmult0\tx886\tx886 \up
0 \expndtw0\charscalex101 100 mL of food, hence benefiting human health (
\ul0\nosupersub\cf12\f13\fs16 Jayamanne & \line \up0 \expndtw0\charscalex103 Ad
ams, 2006\ul0\nosupersub\cf10\f11\fs16 ). In Brazil, the present legislation sta
tes that the min-\line \up0 \expndtw0\charscalex108 imum viable quantity of prob
iotic culture should be between 10\ul0\super\cf13\f14\fs16 8 \line \up0 \expndtw
0\charscalex109 \ul0\nosupersub\cf10\f11\fs16 and 10\ul0\super\cf13\f14\fs16 9\
ul0\nosupersub\cf10\f11\fs16 CFU per daily portion of product and that the prob
iotic \line \up0 \expndtw0\charscalex104 population should be stated on the prod
uct label (\ul0\nosupersub\cf12\f13\fs16 ANVISA, 2008\ul0\nosupersub\cf10\f11\fs
16 ). \line\tab \up0 \expndtw0\charscalex106 Several health benefits are attribu
ted to the ingestion of foods \line \up0 \expndtw0\charscalex108 containing prob
iotic cultures, some of them proven scientifically \line \up0 \expndtw0\charscal
ex108 and others still requiring further studies in humans. Some of the \line \u
p0 \expndtw0\charscalex105 main health benefits related to probiotics are: antimicrobial activ-\line \up0 \expndtw0\charscalex101 ity, prevention and treatment
of diarrhea, relief of symptoms caused \line \up0 \expndtw0\charscalex103 by la
ctose intolerance, anti-mutagenic and anti-carcinogenic activ-\line \up0 \expndt
w0\charscalex109 ities, and stimulation of the immune system (\ul0\nosupersub\c
f12\f13\fs16 Shah, 2007\ul0\nosupersub\cf10\f11\fs16 ). It is \line \up0 \expnd
tw0\charscalex108 important to emphasize that the effects on health promotion ar
e \line \up0 \expndtw0\charscalex103 strain-dependent and cannot be predicted fo
r a determined specie \line \up0 \expndtw0\charscalex102 of microorganisms, and
that there is no single probiotic strain capa-\line \up0 \expndtw0\charscalex102
ble of providing all the benefits mentioned above (\ul0\nosupersub\cf12\f13\fs1
6 Shah, 2007\ul0\nosupersub\cf10\f11\fs16 ). \line\tab \up0 \expndtw0\charscalex
104 The dairy industry, in particular, has found probiotic cultures to \line \up
0 \expndtw0\charscalex106 be a tool for the development of new functional produc
ts (\ul0\nosupersub\cf12\f13\fs16 Cham-\line \up0 \expndtw0\charscalex108 pagne
et al., 2005\ul0\nosupersub\cf10\f11\fs16 ). Yoghurts and fermented milks are th
e main \line \up0 \expndtw0\charscalex107 vehicles for probiotic cultures. Howev
er, new products are being \line \up0 \expndtw0\charscalex109 introduced in the
international market, such as milk-based des-\line \up0 \expndtw0\charscalex103
serts, powdered milk for newborn infants, ice-creams, butter, may-\line \up0 \ex
pndtw0\charscalex103 onnaise, various types of cheese, products in the form of c
apsules \line \up0 \expndtw0\charscalex109 or powders to be dissolved in cold dr
inks, and fermented foods \line \up0 \expndtw0\charscalex106 of vegetable origin
(\ul0\nosupersub\cf12\f13\fs16 Champagne et al., 2005; Komatsu, Buriti, & \li
ne \up0 \expndtw-2\charscalex100 Saad, 2008; Saad, 2006\ul0\nosupersub\cf10\f11\
fs16 ).\par\pard\qj \li654\ri150\sb2\sl-209\slmult0\fi232 \up0 \expndtw0\charsca
lex106 There is thus an increased variety of products available in the \up0 \exp
ndtw0\charscalex103 market and consumers are getting more used to the probiotic
con-\line \up0 \expndtw0\charscalex102 cept, with a consequent increased demand
for such products. Con-\line \up0 \expndtw0\charscalex107 sequently, it turned o
ut to be necessary to acquire knowledge of \up0 \expndtw0\charscalex101 the vari
ous operations currently used in the processing of a specific \up0 \expndtw0\cha
rscalex105 food product, and the level of influence - positive or negative - on
\up0 \expndtw0\charscalex104 the survival of these microorganisms.\par\pard\qj \
li654\ri149\sb1\sl-209\slmult0\fi232 \up0 \expndtw0\charscalex107 The aim of the
present paper was to review the technological \up0 \expndtw0\charscalex104 para
meters involved in the production of probiotic ice-creams. \up0 \expndtw0\
charscalex108 Although the application of probiotics in cheeses and especially \
up0 \expndtw0\charscalex113 in fermented milks has been widely explored in the l
iterature, \up0 \expndtw0\charscalex110 ice-cream is a relatively innovative mat
rix for the application of \up0 \expndtw0\charscalex106 this microbial group, an
d thus a review about its potential as pro-\line \up0 \expndtw0\charscalex106 bi
otic food carrier could be very helpful.\par\pard\ql \li654\sb0\sl-184\slmult0 \
par\pard\ql \li654\sb0\sl-184\slmult0 \par\pard\ql \li654\sb68\sl-184\slmult0 \u
p0 \expndtw0\charscalex108 2. Probiotic ice-cream processing\par\pard\qj \li654\
sb0\sl-209\slmult0 \par\pard\qj \li654\ri149\sb5\sl-209\slmult0\fi232 \up0 \expn
dtw0\charscalex104 Ice-cream is a frozen mixture of a combination of components,
\line \up0 \expndtw0\charscalex103 such as milk, sweeteners, stabilizers,
emulsifiers and flavoring \line \up0 \expndtw0\charscalex101 agents (\ul0\nosu
persub\cf12\f13\fs16 Marshall, Goff, & Hartel, 2003\ul0\nosupersub\cf10\f11\fs16
). This category includes sev-\line \up0 \expndtw0\charscalex102 eral related p
roducts, such as plain ice-cream, reduced fat, low fat, \line \up0 \expndtw0\cha
rscalex107 nonfat, fruit, and nut ice-creams, puddings, variegated, mousse, \lin
e \up0 \expndtw0\charscalex106 sherbet, frozen yoghurt, besides other frozen pro
ducts (\ul0\nosupersub\cf12\f13\fs16 Marshall \line \up0 \expndtw0\charscalex103
& Arbuckle, 1996\ul0\nosupersub\cf10\f11\fs16 ).\par\pard\qj \li654\ri149\sb2\s
l-209\slmult0\fi232 \up0 \expndtw0\charscalex107 Freezing involves vigorously ag
itating to incorporate air, thus \line \up0 \expndtw0\charscalex108 conferring t
he desirable smoothness and softness of the frozen \line \up0 \expndtw0\charscal
ex104 product (\ul0\nosupersub\cf12\f13\fs16 Marshall et al., 2003\ul0\nosupersu
b\cf10\f11\fs16 ). Ice-cream is highly accepted prod-\line \up0 \expndtw0\charsc

alex105 uct by children, adolescents, and adults, as well as by the elderly \lin
e \up0 \expndtw0\charscalex106 public. Due to its rather refreshing features, it
is more consumed \line \up0 \expndtw0\charscalex109 in the summer, but some peo
ple have the habit of consuming it \line \up0 \expndtw0\charscalex104 throughout
the year. In 2003, 5333 million liters of ice-cream were \line \up0 \expndtw0\c
harscalex106 produced in the USA. Therefore, 10% of the total milk production \l
ine \up0 \expndtw0\charscalex102 and 16% of the processed milk were used for thi
s purpose. In Can-\line \up0 \expndtw0\charscalex108 ada, 380 million liters of
milk were used to produce desserts, of \line \up0 \expndtw0\charscalex109 which
79% was used for the production of full-fat or low-fat ice-\line \up0 \expndtw0\
charscalex106 cream (\ul0\nosupersub\cf12\f13\fs16 Goff & Griffiths, 2006\ul0\
nosupersub\cf10\f11\fs16 ). In 2006, although an overall de-\line \up0 \expndtw
0\charscalex104 crease in the product sales was registered in the American super
-\line \up0 \expndtw0\charscalex107 markets, the sale of light and diet ice-crea
ms increased by 15%\par\pard\column \ql \li6034\sb0\sl-184\slmult0 \par\pard\ql
\li20\sb59\sl-184\slmult0 \up0 \expndtw0\charscalex104 from January to June
\up0 \expndtw0\charscalex105 (\ul0\nosupersub\cf12\f13\fs16 International Dairy
Foods Association,\par\pard\qj \li20\ri646\sb4\sl-209\slmult0 \up0 \expndtw0\
charscalex100 2007\ul0\nosupersub\cf10\f11\fs16 ). In Brazil, a 7.6% increa
se in the product sales between \up0 \expndtw0\charscalex105 2005 and 200
6 was observed, when the annual per capita con-\line \up0 \expndtw0\charscalex1
04 sumption varied from 1202 to 1252 L (\ul0\nosupersub\cf12\f13\fs16 Hegg, 2007
\ul0\nosupersub\cf10\f11\fs16 ). These figures \up0 \expndtw0\charscalex105 summ
arize an incredible low consumption for a country with such \up0 \expndtw0\chars
calex105 large area and where the majority of the states present high tem-\line
\up0 \expndtw0\charscalex105 peratures during the whole year.\par\pard\qj \li20\
ri646\sb1\sl-209\slmult0\fi232 \up0 \expndtw0\charscalex104 The ice-cream matrix
might be a good vehicle for probiotic cul-\line \up0 \expndtw0\charscalex104 tu
res, due to its composition, which includes milk proteins, fat and \up0 \expndtw
0\charscalex103 lactose, as well as other compounds. Moreover, the fact that it
is a \up0 \expndtw0\charscalex105 frozen product certainly contributes. However,
the product should \up0 \expndtw0\charscalex105 have relatively high pH values
- from 5.5 up to 6.5, which leads to \up0 \expndtw0\charscalex102 an increased s
urvival of the lactic cultures during storage; also, the \up0 \expndtw0\charscal
ex103 lower acidity results in increased consumer acceptance, especially \up0 \e
xpndtw0\charscalex106 by those who prefer mild products.\par\pard\ql \li20\ri645
\sb2\sl-209\slmult0\fi232\tx262\tx262 \up0 \expndtw0\charscalex102 Even though m
any ice-cream formulations are rich in sugar and \line \up0 \expndtw0\charscalex
106 fat, ice-cream is generally considered as a nutritive food, since it \line \
up0 \expndtw0\charscalex104 contains milk, and sometimes fruit, in its formulati
on. The addition \line \up0 \expndtw0\charscalex107 of probiotic cultures to ice
-creams, in addition to adding value to \line \up0 \expndtw0\charscalex103 the p
roduct, provides it with the advantage of being functional. Be-\line \up0 \expnd
tw0\charscalex107 sides, the regular consumption of probiotic ice-cream containi
ng \line \up0 \expndtw0\charscalex105 Bifidobacterium lactis Bb-12 was reported
to decrease the viable \line \up0 \expndtw0\charscalex106 counts of salivary st
reptococci and lactobacilli, caries-associated \line \up0 \expndtw0\charscalex10
6 microorganisms in the oral cavity, probably due to an adherence \line \up0 \ex
pndtw0\charscalex107 to the oral mucosa and to the dental tissues, as part of a
biofilm, \line \up0 \expndtw0\charscalex103 and therefore competing with oral pa
thogens (\ul0\nosupersub\cf12\f13\fs16 aglar et al., 2008\ul0\nosupersub\cf10\f11
\fs16 ). \line\tab \up0 \expndtw0\charscalex105 During probiotic ice-cream pr
oduction, each process stage \line \up0 \expndtw0\charscalex105 ought to be o
ptimized, aiming at an increased survival of the pro-\line \up0 \expndtw0\charsc
alex107 biotic bacteria, so as to guarantee the product functional proper-\line
\up0 \expndtw0\charscalex110 ties. This means that the main challenges in
volved in the \line \up0 \expndtw0\charscalex106 production of conventional ic
e-cream nowadays must be also ta-\line \up0 \expndtw0\charscalex110 ken into con
sideration during the development of probiotic ice-\line \up0 \expndtw0\charscal
ex105 cream. These challenges include: the contributions for the micro-\line \up
0 \expndtw0\charscalex105 structure and colloidal properties provided by t
he ingredients \line \up0 \expndtw0\charscalex105 and/or components present i
n the formulation; the knowledge \line \up0 \expndtw0\charscalex104 and cont
rol of the ice crystallization; the choice of appropriate sta-\line \up0 \expndt
w0\charscalex103 bilizers and, finally, the understanding and control of the fat
desta-\line \up0 \expndtw0\charscalex107 bilization and the emulsifier function
ality (\ul0\nosupersub\cf12\f13\fs16 Goff, 2008\ul0\nosupersub\cf10\f11\fs16 ).
\line\tab \up0 \expndtw0\charscalex108 In other words, the incorporation of prob
iotic bacteria into an \line \up0 \expndtw0\charscalex108 ice-cream formulation
must not affect the product global quality. \line \up0 \expndtw0\charscalex104 T
herefore, the physical-chemical parameters involved in the qual-\line \up0 \expn
dtw0\charscalex102 ity control of this product, such as the melting rate, and th
e sensory \line \up0 \expndtw0\charscalex102 features, ought to be the same or e
ven better, when compared to a \line \up0 \expndtw0\charscalex103 conventional i
ce-cream.\par\pard\qj \li20\ri646\sb1\sl-209\slmult0\fi232 \up0 \expndtw0\charsc
alex102 Overall, the general steps of probiotic ice-cream processing are: \line
\up0 \expndtw0\charscalex107 reception/weighing of the ingredients involved (mil
k, emulsifiers, \line \up0 \expndtw0\charscalex105 stabilizers, milk powder, sug
ar); mixing; pasteurizing; cooling to a \line \up0 \expndtw0\charscalex102 tempe
rature of around 37-40 C, for the addition of the freeze-dried \line \up0 \expn
dtw0\charscalex102 starter cultures (usually yoghurt cultures) and the probiotic
cultures \line \up0 \expndtw0\charscalex105 (adjunct cultures); subsequent ferm
entation to a pH of 4.8-4.7, or \line \up0 \expndtw0\charscalex108 the addition
of a previously fermented inoculum containing both \line \up0 \expndtw0\charscal
ex105 types of lactic cultures; cooling to 4 \up0 \expndtw0\charscalex104 C and
keeping the mixture at\par\pard\ql \li20\sb22\sl-184\slmult0 \up0 \expndtw0\char
scalex106 this temperature (4 \up0 \expndtw0\charscalex102 C) for 24 h for the m
aturation. The processing\par\pard\qj \li20\ri645\sb4\sl-209\slmult0 \up0 \expnd
tw0\charscalex106 steps up to this point lead to the production of the ice-cream
mix. \up0 \expndtw0\charscalex101 The mix is subsequently beaten/frozen, in ord
er to produce the final \up0 \expndtw0\charscalex103 product, which is packag
ed and maintained frozen throughout \up0 \expndtw0\charscalex106 transport,
commercial distribution and maintenance, and storage \up0 \expndtw0\charscalex10
2 for consumption. During all these steps after freezing, the tempera-\line \up0
\expndtw0\charscalex104 ture of the frozen product should be strictly controlle
d.\par\pard\qj \li20\ri985\sb205\sl-209\slmult0 \up0 \expndtw0\charscalex112 3.
Technological hurdles for the incorporation of probiotic \up0 \expndtw0\charscal
ex110 bacteria into ice-cream\par\pard\qj \li20\ri647\sb210\sl-209\slmult0\fi232
\up0 \expndtw0\charscalex104 The incorporation of probiotic bacteria into ice-c
reams is highly \line \up0 \expndtw0\charscalex103 advantageous since, in additi
on to being a rich food from the nutri-\line \up0 \expndtw0\charscalex109 tional
point of view, containing dairy raw material, vitamins and \par\pard\sect\sectd
\fs24\paperw11900\paperh15860\pard\sb0\sl-240{\bkmkstart Pg3}{\bkmkend Pg3}\par\
pard\li850\sb0\sl-149\slmult0\par\pard\li850\sb0\sl-149\slmult0\par\pard\li850\s
b0\sl-149\slmult0\par\pard\li850\sb0\sl-149\slmult0\par\pard\li850\sb105\sl-149\
slmult0\fi3262\tx10946 \up0 \expndtw0\charscalex101 \ul0\nosupersub\cf7\f8\fs13
A.G. Cruz et al. / Food Research International 42 (2009) 1233-1239\tab \up0 \exp
ndtw0\charscalex101 1235\par\pard\li850\sb0\sl-149\slmult0\par\pard\li850\sb97\s
l-149\slmult0\fi0 \up0 \expndtw0\charscalex101 \ul0\nosupersub\cf19\f20\fs13 Tab
le 1\par\pard\li850\sb23\sl-149\slmult0\fi0 \up0 \expndtw0\charscalex102 Technol
ogical hurdles faced during the processing of ice-cream containing probiotic cul
tures.\par\pard\li850\sb139\sl-149\slmult0\fi0\tx3216\tx7849 \up0 \expndtw0\char
scalex101 \ul0\nosupersub\cf7\f8\fs13 Step\tab \up0 \expndtw0\charscalex101 Prob
lem\tab \up0 \expndtw0\charscalex101 Solutions\par\pard\sect\sectd\sbknone\cols2
\colno1\colw7689\colsr160\colno2\colw3911\colsr160\ql \li850\sb105\sl-149\slmult
0\tx3211 \up0 \expndtw0\charscalex103 Adequate choice of the\tab \up0 \expndtw0\
charscalex106 High acidity of the final product may lead to decreased probiotic\
par\pard\ql \li1031\sb23\sl-149\slmult0\tx3211 \up0 \expndtw0\charscalex107 form
ulation ingredients\tab \up0 \expndtw0\charscalex106 survival, in addition to fr
equently be disadvantageous for its\par\pard\ql \li1031\sb23\sl-149\slmult0\tx32

11 \up0 \expndtw0\charscalex112 (fruit pulp/juice)\tab \up0 \expndtw0\charscalex
102 sensory performance\par\pard\ql \li3216\sb24\sl-149\slmult0 \up0 \expndtw0\c
harscalex106 Some ingredients may have inhibitory activity against probiotic\par
\pard\ql \li3216\sb23\sl-149\slmult0 \up0 \expndtw0\charscalex103 strains\par\pa
rd\ql \li850\sb98\sl-149\slmult0\tx3211 \up0 \expndtw0\charscalex104 Preparation
of the\tab \up0 \expndtw0\charscalex106 Probiotic bacteria show some loss in vi
ability at low pH values\par\pard\ql \li1031\sb23\sl-149\slmult0 \up0 \expndtw0\
charscalex107 fermented inoculum\par\pard\ql \li1031\sb25\sl-149\slmult0 \up0 \e
xpndtw0\charscalex111 with the probiotic\par\pard\ql \li1031\sb23\sl-149\slmult0
\up0 \expndtw0\charscalex105 cultures\par\pard\ql \li850\sb98\sl-149\slmult0\tx
3211 \up0 \expndtw0\charscalex100 Beating\tab \up0 \expndtw0\charscalex103 Oxyge
n represents a factor of toxicity for probiotic bacteria, which\par\pard\ql \li3
216\sb23\sl-149\slmult0 \up0 \expndtw0\charscalex105 present anaerobic and/or mi
cro-aerophilic metabolism\par\pard\ql \li850\sb98\sl-149\slmult0\tx3211 \up0 \ex
pndtw-2\charscalex100 Storage\tab \up0 \expndtw0\charscalex105 Stress induced by
freezing reduces the viability of probiotic\par\pard\ql \li3216\sb24\sl-149\slm
ult0 \up0 \expndtw0\charscalex104 bacteria by at least 1 log cycle\par\pard\colu
mn \ql \li20\sb105\sl-149\slmult0 \up0 \expndtw0\charscalex105 UPreferably, use
of fruit pulp/juices with a lower natural\par\pard\ql \li20\sb23\sl-149\slmult0
\up0 \expndtw0\charscalex108 acidity\par\pard\qj \li7849\sb0\sl-171\slmult0 \par
\pard\qj \li20\ri451\sb5\sl-171\slmult0 \up0 \expndtw0\charscalex107 UCheck for
inhibitory activity of the food ingredients to \line \up0 \expndtw0\charscalex10
5 be used against the probiotic strains to be employed\par\pard\qj \li20\ri661\s
b75\sl-171\slmult0 \up0 \expndtw0\charscalex106 UControl of the pH during the fe
rmentative process \up0 \expndtw0\charscalex106 UIncreased inoculum concentratio
n\par\pard\ql \li20\sb21\sl-149\slmult0 \up0 \expndtw0\charscalex107 USelection
of strains tolerant to low pH values\par\pard\ql \li7849\sb0\sl-149\slmult0 \par
\pard\ql \li20\sb120\sl-149\slmult0 \up0 \expndtw0\charscalex106 USelection of o
xygen-tolerant strains\par\pard\ql \li7849\sb0\sl-149\slmult0 \par\pard\ql \li20
\sb120\sl-149\slmult0 \up0 \expndtw0\charscalex106 UIncreased inoculum concentra
tion\par\pard\qj \li7849\sb0\sl-171\slmult0 \par\pard\qj \li20\ri527\sb6\sl-171\
slmult0 \up0 \expndtw0\charscalex106 UAvoid temperature oscillations during stor
age of the \line \up0 \expndtw0\charscalex107 product\par\pard\sect\sectd\sbknon
e\cols2\colno1\colw6070\colsr160\colno2\colw5530\colsr160\ql \li850\sb0\sl-184\s
lmult0 \par\pard\ql \li850\sb0\sl-184\slmult0 \par\pard\ql \li850\sb0\sl-184\slm
ult0 \par\pard\ql \li850\sb140\sl-184\slmult0 \up0 \expndtw0\charscalex106 \ul0\
nosupersub\cf10\f11\fs16 minerals in its composition, it is usually consumed by
everybody,\par\pard\ql \li850\ri149\sb5\sl-209\slmult0\tx1082\tx1082 \up0 \expnd
tw0\charscalex105 being well accepted by the public. In the specific case of pro
biotic \line \up0 \expndtw0\charscalex105 ice-cream, this is a concrete challeng
e as, most of the times, ice-\line \up0 \expndtw0\charscalex106 cream is not con
sumed daily by most of the consumers, and this \line \up0 \expndtw0\charscalex10
8 frozen dessert is more frequently consumed during the summer \line \up0 \expnd
tw0\charscalex100 in most of the countries, and it is hence considered as an occ
asional \line \up0 \expndtw0\charscalex107 food. Therefore, the launching of ice
-creams supplemented with \line \up0 \expndtw0\charscalex107 probiotic cultures
should be accompanied by educational cam-\line \up0 \expndtw0\charscalex105 pa
igns aiming to encourage consumers towards a more constant \line \up0 \expndtw0\
charscalex104 consumption and showing them the benefits provided by this
\line \up0 \expndtw0\charscalex104 change of behavior. The fact that consumers
are not aware of the \line \up0 \expndtw0\charscalex105 benefits caused by consu
ming foods containing probiotic bacteria \line \up0 \expndtw0\charscalex100 was
recently reported (\ul0\nosupersub\cf12\f13\fs16 Viana, Cruz, Zoellner, Silva, &
Batista, 2008\ul0\nosupersub\cf10\f11\fs16 ). \line\tab \up0 \expndtw0\charscal
ex112 However, the technology used to transform this food into a \line \up0 \exp
ndtw0\charscalex104 vehicle for probiotic microorganisms must be well und
erstood \line \up0 \expndtw0\charscalex104 and studied simultaneously with the k
nowledge of the metabolism \line \up0 \expndtw0\charscalex105 of the added cultu
res. This should be done so as to provide suffi-\line \up0 \expndtw0\charscalex1
11 cient viability of the microorganisms in the product throughout \line \up0 \e

xpndtw0\charscalex105 its shelf life, thus resulting in therapeutic activity for
the consumer, \line \up0 \expndtw0\charscalex106 without any negative influence
on the sensory acceptance of the \line \up0 \expndtw0\charscalex109 product and
on the functional properties. Overall, it is observed \line \up0 \expndtw0\char
scalex106 that the growth and survival of probiotic bacteria is strain-depen-\li
ne \up0 \expndtw0\charscalex102 dent (\ul0\nosupersub\cf12\f13\fs16 Homayouni, E
hsani, Azizi, Razavi, & Yarmand, 2008b\ul0\nosupersub\cf10\f11\fs16 ). \line\tab
\up0 \expndtw0\charscalex105 The technological hurdles which take place in the
existing pro-\line \up0 \expndtw0\charscalex104 cessing steps with respect to th
e development of ice-creams con-\line \up0 \expndtw0\charscalex107 taining probi
otic bacteria should be known and optimized (\ul0\nosupersub\cf12\f13\fs16 Table
\line \up0 \expndtw0\charscalex106 1\ul0\nosupersub\cf10\f11\fs16 ). As a gener
al rule, the addition of probiotics strains into a food \line \up0 \expndtw0\cha
rscalex106 matrix implies the need to assure the viability of the probiotic cul\line \up0 \expndtw0\charscalex109 ture at high levels during the storage period
, without altering its \line \up0 \expndtw0\charscalex102 sensory characteristic
s (\ul0\nosupersub\cf12\f13\fs16 Stanton et al., 2003\ul0\nosupersub\cf10\f11\fs
16 ). In the particular case \line \up0 \expndtw0\charscalex104 of frozen produc
ts like ice-cream, this implies overcoming intrinsic \line \up0 \expndtw0\charsc
alex104 hurdles which take place in the processing of ice-creams, such as \line
\up0 \expndtw0\charscalex108 the beating step, where air is incorporated - known
as overrun -\line \up0 \expndtw0\charscalex108 and storage under freezing tempe
ratures, which affects survival \line \up0 \expndtw0\charscalex106 of the probio
tic microorganisms during storage, and also the way \line \up0 \expndtw0\charsca
lex108 in which the bacterial inoculum is added to the product. In addi-\line \u
p0 \expndtw0\charscalex108 tion, great attention should be given to the correct
choice of the \line \up0 \expndtw0\charscalex110 other ingredients to be used in
the product, especially any fruit \line \up0 \expndtw0\charscalex109 pulp/juice
, which will give the final flavor to the product.\par\pard\ql \li850\sb0\sl-184
\slmult0 \par\pard\ql \li850\sb47\sl-184\slmult0 \up0 \expndtw0\charscalex100 3.
1. Fruit pulp or juice as an ingredient of probiotic ice-creams\par\pard\qj \li8
50\sb0\sl-209\slmult0 \par\pard\qj \li850\ri150\sb5\sl-209\slmult0\fi232 \up0 \e
xpndtw0\charscalex103 Fruits, in the form of juice or pulp, are widely used as f
lavorings \line \up0 \expndtw0\charscalex103 in ice-creams, being commercially a
vailable in the form of pasteur-\line \up0 \expndtw0\charscalex112 ized juices o
r frozen pulps, representing the most well known \line \up0 \expndtw0\charscalex
101 source of flavor for this product (\ul0\nosupersub\cf12\f13\fs16 Papademas &
Bintsis, 2000\ul0\nosupersub\cf10\f11\fs16 ). Fruits\par\pard\column \ql \li623
0\sb0\sl-184\slmult0 \par\pard\ql \li6230\sb0\sl-184\slmult0 \par\pard\ql \li623
0\sb0\sl-184\slmult0 \par\pard\ql \li20\sb141\sl-184\slmult0 \up0 \expndtw0\char
scalex106 or their derivatives with a pronounced acidic character should be\par\
pard\qj \li20\ri449\sb5\sl-209\slmult0 \up0 \expndtw0\charscalex104 avoided in
ice-creams containing probiotic cultures, since this \up0 \expndtw0\charsc
alex109 attribute could influence their sensory acceptance and also de-\line \up
0 \expndtw0\charscalex104 creased the viability of the cultures (\ul0\nosu
persub\cf12\f13\fs16 Fvaro-Trindade, Balieiro, \up0 \expndtw0\charscalex102 Dias
, Sanino, & Boschini, 2007; Fvaro-Trindade, Bernadi, Boldini, \up0 \expndtw0\char
scalex103 & Balieiro, 2006\ul0\nosupersub\cf10\f11\fs16 ) as its addition decrea
se pH values.\par\pard\qj \li20\ri450\sb1\sl-209\slmult0\fi232 \up0 \expndtw0\ch
arscalex104 Acid pH tolerance in probiotic bacteria is strain dependent, and \li
ne \up0 \expndtw0\charscalex103 Bifidobacteria strains are more sensitive than L
actobacillus strains \line \up0 \expndtw0\charscalex102 (\ul0\nosupersub\cf12\f1
3\fs16 Godward et al., 2000\ul0\nosupersub\cf10\f11\fs16 ). Even though this spe
cific mechanism is still \line \up0 \expndtw0\charscalex103 not well established
, the influence of a particular enzyme, H\ul0\super\cf13\f14\fs16 +\ul0\nosupers
ub\cf10\f11\fs16 -ATP-\line \up0 \expndtw0\charscalex103 ase, was reported (\ul
0\nosupersub\cf12\f13\fs16 Lankaputhra, Shah, & Britz, 1996; Takahashi, \line \
up0 \expndtw0\charscalex106 Xiao, Miyaji, & Iwatsuki, 2007\ul0\nosupersub\cf10\f
11\fs16 ). The development of strategies to \line \up0 \expndtw0\charscalex102 e
nhance their survival in an adverse environment is essential. \line \up0

\expndtw0\charscalex108 Firstly, the simplest measure is to select strains resi
stant to low \line \up0 \expndtw0\charscalex109 pH values, which means a continu
ous dialogue with the culture \line \up0 \expndtw0\charscalex110 supplier. Howev
er, it is important to take into consideration the \line \up0 \expndtw0\charscal
ex105 fact that, although the numbers of researches covering the devel-\line \up
0 \expndtw0\charscalex106 opment of probiotic dairy foods supplemented with prob
iotic bac-\line \up0 \expndtw0\charscalex107 teria are increasing, there are sti
ll few strains with proven health \line \up0 \expndtw0\charscalex107 benefits be
ing marketed by each supplier. This finding limits the \line \up0 \expndtw0\char
scalex103 options for commercially available probiotic strains, and the easier \
line \up0 \expndtw0\charscalex104 step is to develop mechanisms of low pH tolera
nce in a laboratory \line \up0 \expndtw0\charscalex106 scale with the probiotic
strains available.\par\pard\qj \li20\ri450\sb2\sl-209\slmult0\fi232 \up0 \expndt
w0\charscalex111 It is also appropriate to remember that in vitro evaluation of
\up0 \expndtw0\charscalex103 probiotic bacteria acid tolerance may not reproduce
a good perfor-\line \up0 \expndtw0\charscalex102 mance of the in vivo behavior,
since poor correlation between them \up0 \expndtw0\charscalex105 was reported (
\ul0\nosupersub\cf12\f13\fs16 Morelli, 2007\ul0\nosupersub\cf10\f11\fs16 ).\par\
pard\qj \li20\ri449\sb2\sl-209\slmult0\fi232 \up0 \expndtw0\charscalex103 Howeve
r, the application of sub-lethal stress conditions, as pro-\line \up0 \expndtw0\
charscalex104 longed exposures of probiotic bacteria to very low pH values (2.0\line \up0 \expndtw0\charscalex106 3.0) for a determined time were reported to p
resent good results, \line \up0 \expndtw0\charscalex102 leading to the generatio
n of a stable and highly acid resistant strain \line \up0 \expndtw0\charscalex10
1 (\ul0\nosupersub\cf12\f13\fs16 Collado & Sanz, 2005; Maus & Ingham, 2003\ul0\n
osupersub\cf10\f11\fs16 ). This treatment also \line \up0 \expndtw0\charscalex10
6 introduces phenotypic changes which improve biological proper-\line \up0 \expn
dtw0\charscalex108 ties, as higher fermentative ability and enzymatic act
ivities, \line \up0 \expndtw0\charscalex105 although an individual evaluation fo
r each strain is needed (\ul0\nosupersub\cf12\f13\fs16 Sanz, \line \up0 \expndtw
0\charscalex102 2007\ul0\nosupersub\cf10\f11\fs16 ). A recent study suggests
the addition of chemical com-\line \up0 \expndtw0\charscalex104 pounds - c
arbonate, and citrate salts - at acceptable levels before \line \up0 \expndtw0\c
harscalex103 or during the incubation, in order to eliminate acidic stress throu
gh \line \up0 \expndtw0\charscalex103 chemical reactions. The resultants product
s would be metabolized \line \up0 \expndtw0\charscalex106 in a subsequent step,
with a consequent provision of a favorable \line \up0 \expndtw0\charscalex103 co
ndition for probiotic bacteria growth (\ul0\nosupersub\cf12\f13\fs16 Zhao & Li,
2008\ul0\nosupersub\cf10\f11\fs16 ). This find-\line \up0 \expndtw0\charscalex10
3 ing deserves more investigation.\par\pard\qj \li20\ri450\sb2\sl-209\slmult0\fi
232 \up0 \expndtw0\charscalex103 In a practical and economic vision, fruits and/
or flavorings addi-\line \up0 \expndtw0\charscalex105 tives with mild features (
low acidity values) ought to be used, and \line \up0 \expndtw0\charscalex105 sho
uld be acquired from processing plants which adopt an assur-\line \up0 \expndtw0
\charscalex102 ance quality system (HACCP) to avoid microbial and chemical con{\shp {\*\shpinst\shpleft850\shptop1652\shpright11251\shpbottom4888\shpfhdr0\shp
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}}\par\pard\sect\sectd\fs24\paperw11900\paperh15860\pard\sb0\sl-240{\bkmkstart P
g4}{\bkmkend Pg4}\par\pard\li654\sb0\sl-149\slmult0\par\pard\li654\sb0\sl-149\sl
mult0\par\pard\li654\sb0\sl-149\slmult0\par\pard\li654\sb0\sl-149\slmult0\par\pa
rd\li654\sb104\sl-149\slmult0\fi0\tx3916 \up0 \expndtw-1\charscalex100 \ul0\nosu
persub\cf7\f8\fs13 1236\tab \up0 \expndtw-1\charscalex100 A.G. Cruz et al. / Foo
d Research International 42 (2009) 1233-1239\par\pard\sect\sectd\sbknone\cols2\c
olno1\colw5874\colsr160\colno2\colw5726\colsr160\ql \li654\sb0\sl-184\slmult0 \p
ar\pard\ql \li654\sb58\sl-184\slmult0 \up0 \expndtw0\charscalex107 \ul0\nosupers
ub\cf10\f11\fs16 tamination, However, it is important to point out that
certain\par\pard\qj \li654\ri150\sb4\sl-209\slmult0 \up0 \expndtw0\charscalex10
7 fruits, like passion fruit, may inhibit probiotic microorganisms, \line
\up0 \expndtw0\charscalex111 resulting in loss of viability in the product, due
the presence of \line \up0 \expndtw0\charscalex110 intrinsic factors linked to t
heir composition (\ul0\nosupersub\cf12\f13\fs16 Buriti, Komatsu, & \line \up0 \
expndtw-2\charscalex100 Saad, 2007\ul0\nosupersub\cf10\f11\fs16 ).\par\pard\ql \
li654\sb0\sl-184\slmult0 \par\pard\ql \li654\sb46\sl-184\slmult0 \up0 \expndtw0\

charscalex100 3.2. Addition of probiotic cultures to ice-cream\par\pard\qj \li65
4\sb0\sl-209\slmult0 \par\pard\qj \li654\ri149\sb6\sl-209\slmult0\fi232 \up0 \ex
pndtw0\charscalex106 Independently of the moment of their addition during
ice-\line \up0 \expndtw0\charscalex107 cream processing, probiotic bacteria pl
ay the role of adjunct cul-\line \up0 \expndtw0\charscalex105 tures, which means
that starter cultures - yogurt cultures, in most \up0 \expndtw0\charscalex108 o
f the cases - will frequently be present. In case fermentation of \up0 \expndtw0
\charscalex106 ice-cream is desirable (frozen yoghurt ice-cream), this co-cultur
e \up0 \expndtw0\charscalex108 (starter plus probiotic) turns out to be more fit
ted to an industrial \up0 \expndtw0\charscalex104 vision, as it decreases fermen
tation time.\par\pard\qj \li654\ri149\sb2\sl-209\slmult0\fi232 \up0 \expndtw0\ch
arscalex104 Cultures may be added to ice-creams in two ways, considering \line \
up0 \expndtw0\charscalex106 that they are of the DVS (Direct Vat Set) type, for
the direct addi-\line \up0 \expndtw0\charscalex106 tion to the product during it
s manufacture: either adding them di-\line \up0 \expndtw0\charscalex108 rectly t
o the pasteurized mix or using the milk as a substrate for \line \up0 \expndtw0\
charscalex111 fermentation, producing, in the latter case, frozen yoghurt ice-\l
ine \up0 \expndtw0\charscalex100 cream. In the second case, the pH must be close
ly controlled during \line \up0 \expndtw0\charscalex105 the fermentative process
from the moment of obtaining the inocu-\line \up0 \expndtw0\charscalex103 lum,
and also the temperature during storage, so that any undesir-\line \up0 \expndtw
0\charscalex108 able reactions do not occur during this period. In addition to t
he \line \up0 \expndtw0\charscalex104 increased sensibility of probiotic microor
ganisms to low pH values \line \up0 \expndtw0\charscalex107 (4.0-4.5), negative
effects on sensory acceptance of the product \line \up0 \expndtw0\charscalex103
may arise. These changes might be undesirable, since ice-creams \line \up0 \expn
dtw0\charscalex109 are not traditionally characterized as high acidic food produ
cts. \line \up0 \expndtw0\charscalex108 One alternative is to stop the fermentat
ion at pH values ranging \line \up0 \expndtw0\charscalex103 from 5.0 to 5.5 (\ul
0\nosupersub\cf12\f13\fs16 Vardar & ksz, 2007\ul0\nosupersub\cf10\f11\fs16 ).\par\
pard\qj \li654\ri76\sb0\sl-209\slmult0\fi232\tx3108 \up0 \expndtw0\charscalex101
Probiotic ice-creams using the fermentation as a regular step for \up0 \expndtw
0\charscalex106 their production are naturally more acid, due to the production
of \up0 \expndtw0\charscalex106 lactic acid during the fermentative process (\ul
0\nosupersub\cf12\f13\fs16 Basyg\u728?it, Kuleasan, & \up0 \expndtw0\charscalex1
02 Karahan, 2006\ul0\nosupersub\cf10\f11\fs16 ). Many lactic acid bacteria show
a stationary phase \up0 \expndtw0\charscalex114 with little growth, but also suf
fer a rapid loss of cell viability if \line \up0 \expndtw0\charscalex106 stored
at temperatures below \tab \up0 \expndtw0\charscalex102 20 C (\ul0\nosupersub\c
f12\f13\fs16 Naidu, Bidlack, & Clemens,\par\pard\qj \li654\ri150\sb1\sl-209\slmu
lt0 \up0 \expndtw0\charscalex106 1999\ul0\nosupersub\cf10\f11\fs16 ). In general
, probiotics are added to yogurts and fermented \up0 \expndtw0\charscalex111 mil
k that are stored at temperatures of 5-10 \up0 \expndtw0\charscalex104 C. Such
products\par\pard\qj \li654\ri150\sb1\sl-209\slmult0 \up0 \expndtw0\charscalex10
5 may be more susceptible to loss of culture viability than ice-\line
\up0 \expndtw0\charscalex108 creams, because of the lower storage temperature of
the latter, \up0 \expndtw0\charscalex108 which may result in cell injury due to
freezing.\par\pard\ql \li654\sb0\sl-184\slmult0 \par\pard\ql \li654\sb47\sl-184
\slmult0 \up0 \expndtw0\charscalex100 3.3. Stability of the probiotic cultures d
uring ice-cream storage\par\pard\qj \li654\sb0\sl-209\slmult0 \par\pard\qj \li65
4\ri150\sb5\sl-209\slmult0\fi232 \up0 \expndtw0\charscalex104 The viability of p
robiotics in a delivery system (food matrix) de-\line \up0 \expndtw0\charscalex1
03 pends, among many other factors, on the strain selected, the inter-\line \up0
\expndtw0\charscalex106 actions between the microbial species present, the prod
uction of \up0 \expndtw0\charscalex104 hydrogen peroxide due to bacterial metabo
lism and the final acid-\line \up0 \expndtw0\charscalex105 ity of the product (\
ul0\nosupersub\cf12\f13\fs16 Vasiljevic & Shah, 2008\ul0\nosupersub\cf10\f11\fs1
6 ).\par\pard\ql \li654\ri76\sb1\sl-209\slmult0\fi232\tx886 \up0 \expndtw0\chars
calex109 Studies carried out in various parts of the world have shown \line \up0
\expndtw0\charscalex108 that probiotic cultures were capable of maintaining the

ir stability \line \up0 \expndtw0\charscalex108 in frozen food products, having
minimum loss of viability.\ul0\nosupersub\cf12\f13\fs16 Akalin \line \up0 \expn
dtw0\charscalex104 and Erisir (2008)\ul0\nosupersub\cf10\f11\fs16 investigated
the effect of the addition of prebiotic \line \up0 \expndtw0\charscalex103 ingre
dients (inulin and oligofructose) on the quality parameters, as \line \up0 \expn
dtw0\charscalex103 well as on the probiotic cultures survival (Lactobacillus aci
dophilus \line \up0 \expndtw0\charscalex102 La-5 and Bifidobacterium animalis Bb
-12). Significant differences in \line \up0 \expndtw0\charscalex101 viscosity of
the samples were observed, besides higher overrun val-\line \up0 \expndtw0\char
scalex102 ues for the ice-cream mix with inulin and also less changes in melt-\l
ine \up0 \expndtw0\charscalex105 ing properties. The prebiotic ice-creams also s
howed to be firmer \line \up0 \expndtw0\charscalex106 during storage. The author
s also reported an increased probiotic \line \up0 \expndtw0\charscalex105 surviv
al during storage, mainly for ice-cream containing oligofruc-\line \up0 \expndtw
0\charscalex104 tose as functional ingredient. They concluded that the h
igher \line \up0 \expndtw0\charscalex103 overrun rate values observed for ice-cr
eam containing inulin might \line \up0 \expndtw0\charscalex105 be a hurdle for p
robiotic survival, mainly B. animalis Bb-12. \line\tab \up0 \expndtw0\charscalex
108 \ul0\nosupersub\cf12\f13\fs16 Basyg\u728?it et al. (2006)\ul0\nosupersub\cf1
0\f11\fs16 investigated the survival rates of human \line \up0 \expndtw0\charsc
alex100 probiotic bacteria (L. acidophilus, Lactobacillus agilis and Lactobacil\par\pard\column \ql \li6034\sb0\sl-184\slmult0 \par\pard\ql \li20\sb59\sl-184\s
lmult0 \up0 \expndtw0\charscalex104 lus rhamnosus) in the production of two diff
erent ice-cream formu-\par\pard\ql \li20\sb25\sl-184\slmult0 \up0 \expndtw0\char
scalex107 lations, containing sucrose and aspartame, during \up0 \expndtw0\
charscalex104 6 months\par\pard\ql \li20\sb24\sl-184\slmult0\tx2979 \up0 \expndt
w0\charscalex103 storage. The samples were stored at\tab \up0 \expndtw0\charscal
ex108 20 C and the viability rate\par\pard\qj \li20\ri645\sb5\sl-209\slmult0 \u
p0 \expndtw0\charscalex106 measured monthly. The probiotic cultures were also su
bmitted to \line \up0 \expndtw0\charscalex103 tests of resistance to bile salts,
antibiotics and to hydrochloric acid, \line \up0 \expndtw0\charscalex113 and we
re shown to be highly resistant to all these conditions. \line \up0 \expndtw0\ch
arscalex112 According to the authors, the results showed that none of the \line
\up0 \expndtw0\charscalex108 ice-cream formulations tested, prepared with the ad
dition of dif-\line \up0 \expndtw0\charscalex103 ferent types of sweeteners, cau
sed undesirable effects on the sur-\line \up0 \expndtw0\charscalex109 vival of t
he probiotic bacteria, and the concentrations remained \line \up0 \expndtw0\char
scalex103 constant during storage.\par\pard\qj \li20\ri646\sb1\sl-209\slmult0\fi
232\tx1385\tx2260 \up0 \expndtw0\charscalex101 \ul0\nosupersub\cf12\f13\fs16 Ala
mprese, Foschino, Rossi, Pompei, and Savani (2002)\ul0\nosupersub\cf10\f11\fs16
carried \up0 \expndtw0\charscalex107 out a study with ice-cream supplement
ed with Lactobacillus \up0 \expndtw0\charscalex105 johnsonii La1, simulating i
ndustrial and retail conditions. Four for-\line \up0 \expndtw0\charscalex108 mul
ations containing different amounts of fat (5% and 10% w/v) \up0 \expndtw0\chars
calex100 and sugar (15% and 22% w/v) were prepared. For each formulation, \up0 \
expndtw0\charscalex109 ice-creams with and without La1 were prepared, and store
d at \line \up0 \expndtw0\charscalex103 temperatures of \tab \up0 \expndtw-1\cha
rscalex100 16 C and \tab \up0 \expndtw-2\charscalex100 28 C. The fresh cells o
f La1 and those\par\pard\qj \li20\ri646\sb1\sl-209\slmult0\fi0 \up0 \expndtw0\ch
arscalex104 sampled after freezing were submitted to preliminary tests o
f \up0 \expndtw0\charscalex101 resistance to bile, antibiotics, SDS and acidity.
The microorganisms \up0 \expndtw0\charscalex106 were shown to be resistant to t
he majority of such stress factors, \up0 \expndtw0\charscalex106 leading the aut
hors to exclude the possibility of finding cells sub-\line \up0 \expndtw0\charsc
alex109 lethally injured during freezing. The authors observed high sur-\line \u
p0 \expndtw0\charscalex108 vival rates of the probiotics during storage of the i
ce-creams for \up0 \expndtw0\charscalex108 8 months, with no decrease in the pop
ulation inoculated initially \up0 \expndtw0\charscalex105 (7 log CFU g\ul0\super
\cf13\f14\fs16 1\ul0\nosupersub\cf10\f11\fs16 ). Also, the addition of the pro
biotic microorganism \up0 \expndtw0\charscalex111 did not cause any modification

in the overrun or any alteration \up0 \expndtw0\charscalex106 in firmness of th
e finished product.\par\pard\qj \li20\ri646\sb0\sl-209\slmult0\fi232 \up0 \expnd
tw0\charscalex104 \ul0\nosupersub\cf12\f13\fs16 Alamprese, Foschino, Rossi, Pomp
ei, and Corti (2005)\ul0\nosupersub\cf10\f11\fs16 carried \line \up0 \expndtw0
\charscalex105 out another experiment under the same conditions tested before, \
line \up0 \expndtw0\charscalex102 but using L. rhamnosus GC. Similar results wer
e obtained with this \line \up0 \expndtw0\charscalex103 microorganism, which pre
sented populations of 10\ul0\super\cf13\f14\fs16 8\ul0\nosupersub\cf10\f11\fs16
CFU g\ul0\super\cf13\f14\fs16 1\ul0\nosupersub\cf10\f11\fs16 after \line \u
p0 \expndtw0\charscalex107 one year of storage, with no influence on the physica
l properties \line \up0 \expndtw0\charscalex104 of the ice-cream.\par\pard\qj \l
i20\ri646\sb1\sl-209\slmult0\fi232\tx4087 \up0 \expndtw0\charscalex103 \ul0\nosu
persub\cf12\f13\fs16 Hekmat and McMahon (1992)\ul0\nosupersub\cf10\f11\fs16 det
ermined the survival of L. aci-\line \up0 \expndtw0\charscalex104 dophilus and B
ifidobacterum bifidum in ice-cream to be used as a \line \up0 \expndtw0\charscal
ex103 probiotic food product. The probiotic ice-cream was prepared after \line \
up0 \expndtw0\charscalex106 the fermentation of the ice-cream base mix by both c
ultures, and \line \up0 \expndtw0\charscalex104 then submitted to frozen sto
rage. The survival of the cultures \line \up0 \expndtw0\charscalex109 and t
he activity of -galactosidase were monitored for a period \line \up0 \expndtw0\ch
arscalex104 of 17 weeks in the product during frozen storage at \tab \up0 \expnd
tw-1\charscalex100 29 C. Bacte-\par\pard\qj \li20\ri647\sb2\sl-208\slmult0\tx43
72 \up0 \expndtw0\charscalex104 rial counts were made immediately after freezing
of the fermented \line \up0 \expndtw0\charscalex108 mix, and counts obtained
were, respectively, \up0 \expndtw-1\charscalex100 1.5 \tab \up0 \expndtw0\cha
rscalex102 10\ul0\super\cf13\f14\fs16 8 \up0 \expndtw0\charscalex101 \ul0\nosupe
rsub\cf10\f11\fs16 and\par\pard\ql \li20\sb22\sl-184\slmult0\tx460 \up0 \expndtw
-1\charscalex100 2.5\tab \up0 \expndtw0\charscalex100 10\ul0\super\cf13\f14\fs16
8\ul0\nosupersub\cf10\f11\fs16 CFU mL\ul0\super\cf13\f14\fs16 1\ul0\nosupers
ub\cf10\f11\fs16 . After seventeen weeks of storage, these popu-\par\pard\ql \li
20\sb24\sl-184\slmult0\tx2226\tx3373 \up0 \expndtw0\charscalex102 lations had de
creased to 4\tab \up0 \expndtw0\charscalex100 10\ul0\super\cf13\f14\fs16 6\ul0\n
osupersub\cf10\f11\fs16 and to 1\tab \up0 \expndtw-1\charscalex100 10\ul0\supe
r\cf13\f14\fs16 7\ul0\nosupersub\cf10\f11\fs16 CFU mL\ul0\super\cf13\f14\fs16
1\ul0\nosupersub\cf10\f11\fs16 , respec-\par\pard\qj \li20\ri647\sb7\sl-208\slm
ult0 \up0 \expndtw0\charscalex104 tively. During the same period, the acti
vity of -galactosidase \up0 \expndtw0\charscalex104 decreased from 1800 to 1300
units mL\ul0\super\cf13\f14\fs16 1\ul0\nosupersub\cf10\f11\fs16 .\par\pard\qj
\li20\ri646\sb2\sl-209\slmult0\fi232 \up0 \expndtw0\charscalex104 Sometimes, mo
re additional efforts might be needed, which in-\line \up0 \expndtw0\charscalex1
02 clude the use of the microencapsulation of cultures and the supple-\line \up0
\expndtw0\charscalex108 mentation with prebiotics (\ul0\nosupersub\cf12\f13\fs
16 Akalin & Erisir, 2008; Akyn, Akyn, & \line \up0 \expndtw0\charscalex102 Kirm
aci, 2007; Godward & Kailasapathy, 2003; Kailapasathy & Sul-\line \up0 \expndtw0
\charscalex100 tana, \up0 \expndtw0\charscalex105 2003\ul0\nosupersub\cf10\f11\f
s16 ). Nevertheless, these formulation improvement in-\par\pard\qj \li20\ri
645\sb2\sl-209\slmult0 \up0 \expndtw0\charscalex108 creases the products\ul0\n
osupersub\cf11\f12\fs16 \u8217?\ul0\nosupersub\cf10\f11\fs16 cost. Also, the
products ought to be \up0 \expndtw0\charscalex109 submitted to a sensory ev
aluation, in order to check the impact \up0 \expndtw0\charscalex102 of microcaps
ules and their sensory and textural features, as some-\line \up0 \expndtw0\chars
calex105 times they might lead to rejection by the consumers (\ul0\nosupersub\cf
12\f13\fs16 Homayouni, \up0 \expndtw0\charscalex100 Azizi, Ehsani, Yarmand, & Ra
zavi, 2008a\ul0\nosupersub\cf10\f11\fs16 ).\par\pard\ql \li6034\sb0\sl-184\slmul
t0 \par\pard\ql \li20\sb47\sl-184\slmult0 \up0 \expndtw0\charscalex100 3.4. Over
run and probiotic microorganisms\par\pard\qj \li6034\sb0\sl-209\slmult0 \par\par
d\qj \li20\ri646\sb5\sl-209\slmult0\fi232 \up0 \expndtw0\charscalex106 Overrun i
s defined as the ice-cream percentage of volume in-\line \up0 \expndtw0\charscal
ex106 crease in relation to the liquid mix used to make it, and is related \line
\up0 \expndtw0\charscalex101 to the amount of air incorporated during the manuf

acturing process. \line \up0 \expndtw0\charscalex100 This feature defines the st
ructure of the final product, since the pres-\line \up0 \expndtw0\charscalex102
ence of air gives the ice-cream an agreeable light texture and influ-\line \up0
\expndtw0\charscalex105 ences the physical properties of melting and hardness of
the final \par\pard\sect\sectd\fs24\paperw11900\paperh15860\pard\sb0\sl-240{\bk
mkstart Pg5}{\bkmkend Pg5}\par\pard\li850\sb0\sl-149\slmult0\par\pard\li850\sb0\
sl-149\slmult0\par\pard\li850\sb0\sl-149\slmult0\par\pard\li850\sb0\sl-149\slmul
t0\par\pard\li850\sb104\sl-149\slmult0\fi3262\tx10946 \up0 \expndtw-1\charscalex
100 \ul0\nosupersub\cf7\f8\fs13 A.G. Cruz et al. / Food Research International 4
2 (2009) 1233-1239\tab \up0 \expndtw-1\charscalex100 1237\par\pard\sect\sectd\sb
knone\cols2\colno1\colw6070\colsr160\colno2\colw5530\colsr160\ql \li850\sb0\sl-1
84\slmult0 \par\pard\ql \li850\sb58\sl-184\slmult0 \up0 \expndtw0\charscalex105
\ul0\nosupersub\cf10\f11\fs16 product (\ul0\nosupersub\cf12\f13\fs16 Sofjan & Ha
rtel, 2004\ul0\nosupersub\cf10\f11\fs16 ). In addition, it results in the partia
l\par\pard\ql \li850\ri149\sb4\sl-209\slmult0\fi0\tx1082 \up0 \expndtw0\charscal
ex106 coalescence and destabilization of the fat present in the mixture, \line \
up0 \expndtw0\charscalex106 with the formation of an internal lipid structure, c
apable of impris-\line \up0 \expndtw0\charscalex100 oning air bubbles (\ul0\nosu
persub\cf12\f13\fs16 Bolliger, Goff, & Tharp, 2000\ul0\nosupersub\cf10\f11\fs16
). The Brazilian legis-\line \up0 \expndtw0\charscalex106 lation stipulates the
maximum degree of overrun that ice-creams \line \up0 \expndtw0\charscalex102 can
show as 450 g L\ul0\super\cf13\f14\fs16 1\ul0\nosupersub\cf10\f11\fs16 , inde
pendently of the type of ice-cream, and \line \up0 \expndtw0\charscalex100 refer
s to this parameter as apparent density (\ul0\nosupersub\cf12\f13\fs16 Brasil, 1
999\ul0\nosupersub\cf10\f11\fs16 ). The melt-\line \up0 \expndtw0\charscalex109
ing time determined in the melting test is related to the product \line \up0 \ex
pndtw0\charscalex104 structure stability after the overrun, which, for its part,
is influenced \line \up0 \expndtw0\charscalex100 by the type of emulsifier used
in the process. It thus indicates the ex-\line \up0 \expndtw0\charscalex107 ten
t of stabilization and partial coalescence occurring during the \line \up0 \expn
dtw0\charscalex101 product manufacture (\ul0\nosupersub\cf12\f13\fs16 Correia, P
edrini, & Magalhes, 2007\ul0\nosupersub\cf10\f11\fs16 ). \line\tab \up0 \expndtw0
\charscalex104 Oxygen tolerance in probiotic bacteria is also strain-dependent \
line \up0 \expndtw0\charscalex100 (\ul0\nosupersub\cf12\f13\fs16 Kawasaki, Mimur
a, Satoh, Takeda, & Nimura, 2006\ul0\nosupersub\cf10\f11\fs16 ), and bifidobac-\
line \up0 \expndtw0\charscalex104 teria strains are more susceptible than L. aci
dophilus (\ul0\nosupersub\cf12\f13\fs16 Vasiljevic & \line \up0 \expndtw0\charsc
alex103 Shah, 2008\ul0\nosupersub\cf10\f11\fs16 ). Exposure to oxygen during man
ufacture and storage \line \up0 \expndtw0\charscalex108 of dairy products is hig
hly significant for probiotic bacteria. Most \line \up0 \expndtw0\charscalex101
of the Lactobacillus and Bifidobacterium spp. are gut-derived organ-\line \up0 \
expndtw0\charscalex102 isms and they are microaerophilic and anaerobic, respecti
vely, and \line \up0 \expndtw0\charscalex102 therefore unable to synthesize ATP
by respiratory means, depend-\line \up0 \expndtw0\charscalex110 ing strictly on
a fermentative mode of metabolism (\ul0\nosupersub\cf12\f13\fs16 Talwalkar & \li
ne \up0 \expndtw0\charscalex100 Kailasapathy, 2004b\ul0\nosupersub\cf10\f11\fs1
6 ). These microorganisms oxygen-scavenging \line \up0 \expndtw0\charscalex10
4 system is reduced or completely absent, which results in the \line \u
p0 \expndtw0\charscalex103 incomplete reduction of oxygen to hydrogen pero
xide. Conse-\line \up0 \expndtw0\charscalex108 quently, accumulation of toxic o
xygen metabolites occurs - O\ul0\sub\cf20\f21\fs16 2\ul0\nosupersub\cf10\f11\fs
16 , \line \up0 \expndtw0\charscalex107 OH and H\ul0\sub\cf13\f14\fs17 2\ul0
\nosupersub\cf10\f11\fs16 O\ul0\sub\cf13\f14\fs17 2\ul0\nosupersub\cf10\f11\fs16
- in the cell, eventually leading to its death \line \up0 \expndtw0\
charscalex103 (\ul0\nosupersub\cf12\f13\fs16 Champagne et al., 2005; Talwalkar &
Kailasapathy, 2004a; Vasilj-\line \up0 \expndtw0\charscalex101 evic & Shah, 200
8\ul0\nosupersub\cf10\f11\fs16 ). High levels of intracellular H\ul0\sub\cf13\f1
4\fs17 2\ul0\nosupersub\cf10\f11\fs16 O\ul0\sub\cf13\f14\fs17 2\ul0\nosupersub\c
f10\f11\fs16 block fructose-\line \up0 \expndtw0\charscalex102 6-phosphofructo
ketolase, a key enzyme in the sugar metabolism of \line \up0 \expndtw0\charscale

x102 bifidobacteria (\ul0\nosupersub\cf12\f13\fs16 Shah, 1997\ul0\nosupersub\cf1
0\f11\fs16 ). A correlation between the levels of two \line \up0 \expndtw0\chars
calex106 enzymes - NAD-oxidase and NAD-peroxidase - and the oxygen \line \up0
\expndtw0\charscalex108 susceptibility of bifidobacteria was reported; the forme
r enzyme \line \up0 \expndtw0\charscalex102 gives rise to H\ul0\sub\cf13\f14\fs1
7 2\ul0\nosupersub\cf10\f11\fs16 O\ul0\sub\cf13\f14\fs17 2\ul0\nosupersub\cf10\f
11\fs16 , prompting the latter to scavenge this compound \line \up0 \expndtw0\ch
arscalex103 and prevent cell death (\ul0\nosupersub\cf12\f13\fs16 Roy, 2005\ul0\
nosupersub\cf10\f11\fs16 ).\par\pard\qj \li850\ri149\sb1\sl-209\slmult0\fi232 \u
p0 \expndtw0\charscalex102 The selection of oxygen-resistant strains is essentia
l to succeed \up0 \expndtw0\charscalex108 in maintaining the viability of the cu
lture in probiotic ice-creams, \up0 \expndtw0\charscalex109 since the overrun op
eration cannot be eliminated from the pro-\line \up0 \expndtw0\charscalex106 ces
sing procedure.\ul0\nosupersub\cf12\f13\fs16 Salen, Fathi, and Awad (2005)\ul0
\nosupersub\cf10\f11\fs16 reported that \up0 \expndtw0\charscalex103 the differ
ences in the overrun step of probiotic ice-creams were re-\line \up0 \expndtw0\c
harscalex106 lated to the acidifying capacity of the probiotic cultures, which a
f-\line \up0 \expndtw0\charscalex109 fects the nature of the proteins, causing d
enaturation, and also \up0 \expndtw0\charscalex105 affects the product freezing
point.\par\pard\qj \li850\ri149\sb0\sl-209\slmult0\fi232 \up0 \expndtw0\charscal
ex104 A potential alternative to be considered when dealing with \line \
up0 \expndtw0\charscalex106 this inherent hurdle is to microencapsulate the prob
iotic bacteria, \line \up0 \expndtw0\charscalex104 so as to overcome their
oxygen sensitivity and extended the \line \up0 \expndtw0\charscalex109 produ
ct shelf-life. Microencapsulation is a process in which the \line \up0 \expndtw0
\charscalex109 cells are retained within an encapsulating membrane to reduce \li
ne \up0 \expndtw0\charscalex105 cell injury or cell loss (\ul0\nosupersub\cf12
\f13\fs16 Krasaekoopt, Bhandari, & Deeth, 2003\ul0\nosupersub\cf10\f11\fs16 )
. \line \up0 \expndtw0\charscalex107 \ul0\nosupersub\cf12\f13\fs16 Homayouni et
al. (2008a)\ul0\nosupersub\cf10\f11\fs16 investigated the effect of microencap\line \up0 \expndtw0\charscalex110 sulation on the survival of two probiotic str
ains added in a syn-\line \up0 \expndtw0\charscalex106 biotic ice-cream. Two typ
es of synbiotic ice-cream containing 1% \line \up0 \expndtw0\charscalex105 of re
sistant starch with free and encapsulated Lactobacillus casei \line \up0 \expndt
w0\charscalex108 (Lc-01) and B. lactis (Bb-12) were manufactured. Their surviva
l \line \up0 \expndtw0\charscalex110 was monitored during the product stora
ge for \up0 \expndtw0\charscalex104 180 days at\par\pard\ql \li850\sb22\sl-184
\slmult0\tx4049\tx5210 \up0 \expndtw0\charscalex104 20 C. The probiotic viab
ility was 5.1\tab \up0 \expndtw-2\charscalex100 10\ul0\super\cf13\f14\fs16 9\u
l0\nosupersub\cf10\f11\fs16
and 4.1\tab \up0 \expndtw-3\charscalex100 10\ul0
\super\cf13\f14\fs16 9\ul0\nosupersub\cf10\f11\fs16 CFU/\par\pard\ql \li850\sb2
4\sl-184\slmult0\tx3228\tx3627\tx4049\tx4485 \up0 \expndtw0\charscalex113 mL at
day one, and \up0 \expndtw-1\charscalex100 4.2\tab \up0 \expndtw0\charscalex
102 10\ul0\super\cf13\f14\fs16 6\tab \up0 \expndtw0\charscalex101 \ul0\nosupersu
b\cf10\f11\fs16 and\tab \up0 \expndtw-1\charscalex100 1.1\tab \up0 \expndtw0\cha
rscalex103 10\ul0\super\cf13\f14\fs16 7\ul0\nosupersub\cf10\f11\fs16 CFU/mL af
ter\par\pard\ql \li850\sb26\sl-184\slmult0 \up0 \expndtw0\charscalex105 180 days
of storage at 20 \up0 \expndtw0\charscalex110 C, respectively, to the former a
nd to\par\pard\qj \li850\ri149\sb5\sl-209\slmult0 \up0 \expndtw0\charscalex106 t
he latter in the free state in the ice-cream mixture. The microen-\line \up0 \ex
pndtw0\charscalex104 capsulation at probiotic bacteria in calcium alginate
beads in-\line \up0 \expndtw0\charscalex107 creased their survival rate about
30% during the same period of \line \up0 \expndtw0\charscalex105 storage at the
same temperature. According to the authors, these \line \up0 \expndtw0\charscal
ex107 findings reinforce the encapsulation as a useful alternative to in-\line \
up0 \expndtw0\charscalex104 crease the survival rate of probiotic bacteria
in ice-cream. No \line \up0 \expndtw0\charscalex106 significant effect on
the sensory properties of non-fermented \line \up0 \expndtw0\charscalex107
ice-cream in which the resistant starch was added as a prebiotic \line \up0 \exp
ndtw0\charscalex103 compound was observed.\par\pard\column \ql \li6230\sb0\sl-18

4\slmult0 \par\pard\ql \li20\sb58\sl-184\slmult0 \up0 \expndtw0\charscalex100 3.
5. Influence of storage conditions on the characteristics of probiotic\par\pard\
ql \li20\sb25\sl-184\slmult0 \up0 \expndtw-3\charscalex100 ice-creams\par\pard\q
j \li6230\sb0\sl-209\slmult0 \par\pard\qj \li20\ri450\sb5\sl-209\slmult0\fi232 \
up0 \expndtw0\charscalex105 The development and viability of probiotic microorga
nisms, as \up0 \expndtw0\charscalex110 for other microorganisms, are influenced
by the environmental \up0 \expndtw0\charscalex103 temperature around them. They
can be lethally injured by damage \up0 \expndtw0\charscalex105 to their cell wal
ls or rupture of their membranes caused by the ice \up0 \expndtw0\charscalex103
formed in the external medium, or the cells may be ruptured by ice \up0 \expndtw
0\charscalex104 formed inside the cell and by concentration of solutes in the ex
tra-\line \up0 \expndtw0\charscalex106 cellular medium (\ul0\nosupersub\cf12\f13
\fs16 Gill, 2006\ul0\nosupersub\cf10\f11\fs16 ).\par\pard\qj \li20\ri450\sb2\sl209\slmult0\fi232 \up0 \expndtw0\charscalex104 The velocity of dehydratation of
microbial cells depends on the \up0 \expndtw0\charscalex104 permeability of the
cell membrane and on the surface area in rela-\line \up0 \expndtw0\charscalex102
tion to its volume, which depends on the cell shape and size. How-\line \up0 \e
xpndtw0\charscalex102 ever, the size of the ice crystals decreases with increasi
ng freezing \up0 \expndtw0\charscalex103 rates, and larger intracellular ice cry
stals cause greater damage to \up0 \expndtw0\charscalex104 the cells. (\ul0\nosu
persub\cf12\f13\fs16 Gill, 2006\ul0\nosupersub\cf10\f11\fs16 ).\par\pard\ql \li2
0\ri449\sb0\sl-209\slmult0\fi232\tx262 \up0 \expndtw0\charscalex102 Ice-creams c
ontaining probiotic cultures are frozen foods. Thus, \line \up0 \expndtw0\charsc
alex105 rapid freezing of the mix obtained after inoculating with the micro-\lin
e \up0 \expndtw0\charscalex105 organisms, associated to a rigorous temperature c
ontrol through-\line \up0 \expndtw0\charscalex109 out storage of the product, co
ntributes with the maintenance of \line \up0 \expndtw0\charscalex103 the popula
tions of these microorganisms in the recommended \line \up0 \expndtw0\char
scalex104 doses, capable of providing therapeutic and/or preventive activity. \l
ine \up0 \expndtw0\charscalex109 Preliminary tests should be carried out before
and immediately \line \up0 \expndtw0\charscalex103 after freezing of the product
, so as to have an idea of the inoculum \line \up0 \expndtw0\charscalex108 level
of probiotic cultures to be added, and also of their survival \line \up0 \expnd
tw0\charscalex107 rate, consequently arriving at an optimized level of probiotic
cul-\line \up0 \expndtw0\charscalex110 tures to be added to the product during
its manufacture. Some \line \up0 \expndtw0\charscalex112 studies reported that t
he transformation of mix into ice-cream \line \up0 \expndtw0\charscalex103 influ
enced on the viability of the probiotic cultures (\ul0\nosupersub\cf12\f13\fs16
Haynes & Play-\line \up0 \expndtw0\charscalex100 ne, 2002; Magarios, Selaive, Cos
ta, Flores, & Pizarro, 2007\ul0\nosupersub\cf10\f11\fs16 ). \line\tab \up0 \expn
dtw0\charscalex107 In summary, the decline in bacterial populations during freez
-\line \up0 \expndtw0\charscalex101 ing is caused by injury as a result of the p
rocess to which they were \line \up0 \expndtw0\charscalex104 submitted, causing
their death. In addition, mechanical stress \line \up0 \expndtw0\charscal
ex107 caused by agitation and the incorporation of air might result in a \line \
up0 \expndtw0\charscalex105 smaller population of viable cells (\ul0\nosupersub\
cf12\f13\fs16 Akin, 2005\ul0\nosupersub\cf10\f11\fs16 ).\par\pard\ql \li6230\sb0
\sl-184\slmult0 \par\pard\ql \li6230\sb0\sl-184\slmult0 \par\pard\ql \li20\sb68\
sl-184\slmult0 \up0 \expndtw0\charscalex108 4. Sensory features of probiotic ice
-creams\par\pard\qj \li6230\sb0\sl-209\slmult0 \par\pard\qj \li20\ri449\sb5\sl-2
09\slmult0\fi232 \up0 \expndtw0\charscalex103 \ul0\nosupersub\cf12\f13\fs16 Davi
dson, Duncan, Hackney, Eigel, and Boling (2000)\ul0\nosupersub\cf10\f11\fs16 re
ported \line \up0 \expndtw0\charscalex104 that, in general, ice-creams produced
with probiotic cultures pres-\line \up0 \expndtw0\charscalex105 ent a less inten
se aroma and yoghurt flavor than the product pre-\line \up0 \expndtw0\charscalex
109 pared with traditional cultures. Thus, the production of probiotic \line \up
0 \expndtw0\charscalex104 ice-creams with high sensory acceptance is a difficult
task, requir-\line \up0 \expndtw0\charscalex108 ing technical knowledge by the
processors, as sometimes func-\line \up0 \expndtw0\charscalex109 tional ice-cre
am presents poor sensory performance, when \line \up0 \expndtw0\charscalex1

07 compared to a conventional ice-cream \up0 \expndtw0\charscalex104 (\ul0\n
osupersub\cf12\f13\fs16 Aryana & Summers,\par\pard\qj \li20\ri449\sb1\sl-209\s
lmult0 \up0 \expndtw0\charscalex106 2006; Fvaro-Trindade et al., 2006; Hekmat & M
cMahon, 1992\ul0\nosupersub\cf10\f11\fs16 ). \up0 \expndtw0\charscalex107 Howeve
r, it is possible to develop probiotic ice-cream with good \up0 \expndtw0\charsc
alex105 sensorial quality (\ul0\nosupersub\cf12\f13\fs16 Christianesen, Edelten
, Kristiansen, & Nielsen, \up0 \expndtw0\charscalex102 1996; Vardar & ksz, 2007\ul
0\nosupersub\cf10\f11\fs16 ).\par\pard\qj \li20\ri450\sb1\sl-209\slmult0\fi232 \
up0 \expndtw0\charscalex107 Probiotic cultures do not modify the sensory propert
ies of the \up0 \expndtw0\charscalex108 products to which they are added intense
ly (\ul0\nosupersub\cf12\f13\fs16 Champagne et al., \up0 \expndtw0\charscalex10
0 2005; Saxelin et al., 1999\ul0\nosupersub\cf10\f11\fs16 ). L. acidophilus, as
well as the microorgan-\line \up0 \expndtw0\charscalex104 isms used as starter c
ultures in yoghurt manufacture - Lactobacil-\line \up0 \expndtw0\charscalex100 l
us delbrueckii ssp. bulgaricus and Streptococcus thermophilus - are \up0 \expndt
w0\charscalex107 homofermentative and produce lactic acid from lactose metabo-\l
ine \up0 \expndtw0\charscalex107 lism, without gas production.\par\pard\qj \li20
\ri450\sb2\sl-209\slmult0\fi232 \up0 \expndtw0\charscalex105 Bifidobacteria prod
uce acetic and lactic acids in the proportion \line \up0 \expndtw0\charscalex102
of 3:2. The flavor and aroma of acetic acid provide extremely unde-\line \up0 \
expndtw0\charscalex108 sirable off-flavors to dairy products and might require t
he use of \line \up0 \expndtw0\charscalex105 flavoring agents, so as to minimize
or mask this defect, known as \line \up0 \expndtw0\charscalex107 \ul0\nosupersu
b\cf11\f12\fs16 \u8216?\u8216?\ul0\nosupersub\cf10\f11\fs16 probiotic flavor\u82
21?. An innovative and effective alternative to over-\line \up0 \expndtw-1\chars
calex100 come this possible undesired consequences caused by the presence \line
\up0 \expndtw0\charscalex104 of these cultures is the addition of microencapsula
ted cells of pro-\line \up0 \expndtw0\charscalex106 biotic cultures to food prod
ucts such as yoghurts (\ul0\nosupersub\cf12\f13\fs16 Arai, Sakaki, & \par\pard\s
ect\sectd\fs24\paperw11900\paperh15860\pard\sb0\sl-240{\bkmkstart Pg6}{\bkmkend
Pg6}\par\pard\li653\sb0\sl-149\slmult0\par\pard\li653\sb0\sl-149\slmult0\par\par
d\li653\sb0\sl-149\slmult0\par\pard\li653\sb0\sl-149\slmult0\par\pard\li653\sb10
5\sl-149\slmult0\fi0\tx3915 \up0 \expndtw-1\charscalex100 \ul0\nosupersub\cf7\f8
\fs13 1238\tab \up0 \expndtw-1\charscalex100 A.G. Cruz et al. / Food Research In
ternational 42 (2009) 1233-1239\par\pard\sect\sectd\sbknone\cols2\colno1\colw587
3\colsr160\colno2\colw5727\colsr160\ql \li654\sb0\sl-184\slmult0 \par\pard\ql \l
i654\sb57\sl-184\slmult0 \up0 \expndtw0\charscalex109 \ul0\nosupersub\cf12\f13\f
s16 Sugimoto, 1996\ul0\nosupersub\cf10\f11\fs16 ). Nevertheless, it is importan
t to point out that\par\pard\qj \li654\ri148\sb4\sl-209\slmult0 \up0 \expndtw0\c
harscalex107 products like non-fermented probiotic ice-cream do not normally \up
0 \expndtw0\charscalex107 present problems resulting from the microbial metaboli
sm, since \up0 \expndtw0\charscalex109 they are stored at very low temperatures
(< \up0 \expndtw0\charscalex108 18 C), minimizing\par\pard\ql \li654\ri148\sb2\
sl-209\slmult0\tx886 \up0 \expndtw0\charscalex105 the probiotic microorganisms b
iochemical reactions. \line\tab \up0 \expndtw0\charscalex105 Consumers are gener
ally used to products fermented with yo-\line \up0 \expndtw0\charscalex108 ghurt
cultures, in such a way that the aroma and flavor of these \line \up0 \expndtw0
\charscalex108 products are familiar to them. Considering that the probiotic cul
-\line \up0 \expndtw0\charscalex108 tures use different metabolic pathways than
the classic cultures \line \up0 \expndtw0\charscalex109 employed to ferment milk
, the consumers may find the flavor of \line \up0 \expndtw0\charscalex109 the pr
oducts prepared with the addition of these cultures rather \line \up0 \expndtw0\
charscalex102 unusual.\ul0\nosupersub\cf12\f13\fs16 Vardar and ksz (2007)\ul0\nos
upersub\cf10\f11\fs16 reported that artisan strawberry \line \up0 \expndtw0\cha
rscalex103 ice-cream supplemented with L. acidophilus showed good sensory \line
\up0 \expndtw0\charscalex101 acceptance and that the incubation of the mix at pH
5.6 lead to bet-\line \up0 \expndtw0\charscalex103 ter results in terms of flav
or and taste. The authors suggested that \line \up0 \expndtw0\charscalex106 addi
ng highly acidic fruit to ice-cream might be useful in masking \line \up0 \expnd
tw0\charscalex105 the sour taste resulting from the metabolism of probiotic cult
ures. \line \up0 \expndtw0\charscalex101 However, the results should have not ge
neralized, as opposite find-\line \up0 \expndtw0\charscalex106 ings were reporte
d by\ul0\nosupersub\cf12\f13\fs16 Fvaro-Trindade et al. (2006)\ul0\nosupersub\cf
10\f11\fs16 for probiotic \line \up0 \expndtw0\charscalex107 ice-cream with ace
rola fruit.\par\pard\qj \li654\ri149\sb1\sl-209\slmult0\fi232 \up0 \expndtw0\cha
rscalex111 Therefore, preliminary tests with different inoculation levels \up0 \
expndtw0\charscalex104 and different probiotic microorganisms are recommended in
order \up0 \expndtw0\charscalex106 to optimize the sensory properties of the pr
oduct, mainly with re-\line \up0 \expndtw0\charscalex104 spect to the product ac
idity and sensory performance.\par\pard\ql \li654\sb0\sl-184\slmult0 \par\pard\q
l \li654\sb81\sl-184\slmult0 \up0 \expndtw0\charscalex103 5. Perspectives\par\pa
rd\ql \li654\sb0\sl-209\slmult0 \par\pard\ql \li654\ri148\sb5\sl-209\slmult0\fi2
32\tx886 \up0 \expndtw0\charscalex102 Ice-creams are food products showing great
potential for use as \line \up0 \expndtw0\charscalex106 vehicles for probiotic
cultures, with the advantage of being foods \line \up0 \expndtw0\charscalex103 c
onsumed by all age groups. However, several factors in their pro-\line \up0 \exp
ndtw0\charscalex111 cessing stages ought to be optimized, in order to maintain t
he \line \up0 \expndtw0\charscalex106 microorganisms in viable doses capable of
providing therapeutic \line \up0 \expndtw0\charscalex103 activity to consumers.
In order to achieve this goal, several factors \line \up0 \expndtw0\charscalex10
8 should be strictly controlled, including: the appropriate selection \line \up0
\expndtw0\charscalex103 of cultures to be used, the inoculum concentration, the
appropriate \line \up0 \expndtw0\charscalex102 processing stage for the culture
s to be added, and the strict control \line \up0 \expndtw0\charscalex102 of the
processing procedures and of the transport and storage tem-\line \up0 \expndtw0\
charscalex105 peratures. In conclusion, probiotic cultures do not modify the sen
-\line \up0 \expndtw0\charscalex104 sory features of ice-creams and frozen
desserts strongly and \line \up0 \expndtw0\charscalex105 normally present goo
d viability during the product storage period. \line\tab \up0 \expndtw0\charscal
ex106 Even though several studies have shown adequate viability of \line \up0 \e
xpndtw0\charscalex107 the probiotic cultures during storage of ice-creams, more
clinical \line \up0 \expndtw0\charscalex109 studies on the consumption of probio
tic ice-creams are recom-\line \up0 \expndtw0\charscalex105 mended. Also, it is
important to confirm if, after long storage peri-\line \up0 \expndtw0\charscalex
106 ods, the probiotic cultures are still able to confer the same health \line \
up0 \expndtw0\charscalex108 benefits already observed in other foods with shorte
r shelf-lives \line \up0 \expndtw0\charscalex103 and higher storage temperatures
, such as yoghurts and fermented \line \up0 \expndtw0\charscalex106 milks. Moreo
ver, the launching of ice-creams supplemented with \line \up0 \expndtw0\charscal
ex107 probiotic cultures should be accompanied by educational cam-\line \up0 \
expndtw0\charscalex105 paigns aiming to encourage consumers towards a more const
ant \line \up0 \expndtw0\charscalex104 consumption and showing them the ben
efits provided by this \line \up0 \expndtw0\charscalex102 change of behavior.
\par\pard\ql \li654\sb0\sl-184\slmult0 \par\pard\ql \li654\sb103\sl-184\slmult0
\up0 \expndtw0\charscalex108 Acknowledgements\par\pard\qj \li654\sb0\sl-209\slmu
lt0 \par\pard\qj \li654\ri149\sb5\sl-209\slmult0\fi239 \up0 \expndtw-1\charscale
x100 The authors thank to Fundao de Amparo Pesquisa do Estado \up0 \expndtw0\chars
calex100 de So Paulo (FAPESP), Coordenao de Aperfeioamento de Pes-\line \up0 \expndt
w0\charscalex101 soal de Nvel Superior (CAPES), and Conselho Nacional de Desen-\l
ine \up0 \expndtw0\charscalex106 volvimento Cientfico e Tecnolgico (CNPq) for fin
ancial support \up0 \expndtw0\charscalex101 and scholarships.\par\pard\ql \li654
\sb0\sl-184\slmult0 \par\pard\ql \li654\sb46\sl-184\slmult0 \up0 \expndtw0\chars
calex101 References\par\pard\ql \li654\sb0\sl-159\slmult0 \par\pard\ql \li654\ri
149\sb38\sl-159\slmult0\tx893\tx893 \up0 \expndtw0\charscalex102 \ul0\nosupersub
\cf7\f8\fs13 Akalin, A. S., & Erisir, D. (2008). Effects of inulin and oligofruc
tose on the rheological \line\tab \up0 \expndtw0\charscalex111 characteristics a
nd probiotic culture survival in low-fat probiotic ice-cream. \line \tab \up0 \e
xpndtw-1\charscalex100 Journal of Food Science, 73(4), 184-188.\par\pard\column
\ql \li6034\sb0\sl-149\slmult0 \par\pard\ql \li21\sb97\sl-149\slmult0 \up0 \expn
dtw0\charscalex103 Akin, M. S. (2005). Effects of inulin and different sugar lev

els on viability of probiotic\par\pard\qj \li259\ri646\sb4\sl-158\slmult0 \up0 \
expndtw0\charscalex103 bacteria and the physical and sensory characteristics of
probiotic fermented ice-\line \up0 \expndtw0\charscalex101 cream. Milchwissensch
aft, 60(3), 297-300.\par\pard\ql \li21\ri646\sb2\sl-159\slmult0\tx269\tx269\tx26
9\tx269 \up0 \expndtw0\charscalex100 Akyn, M. B., Akyn, M. S., & Kirmaci, Z. (20
07). Effects of inulin and sugar levels on the \line\tab \up0 \expndtw0\charscal
ex107 viability of yoghurt and probiotic bacteria and the physical and
sensory \line \tab \up0 \expndtw0\charscalex103 characteristics in probiotic ic
e-cream. Food Chemistry, 104(1), 93-99. \line \up0 \expndtw-2\charscalex100 Alam
prese, C., Foschino, R., Rossi, M., Pompei, C., & Corti, S. (2005).
Effects of \line\tab \up0 \expndtw0\charscalex102 Lactobacillus rhamnosus CG ad
dition in ice-cream. International Journal of Dairy \line \tab \up0 \expndtw0\ch
arscalex100 Technology, 58(4), 200-206.\par\pard\qj \li21\ri646\sb0\sl-159\slmul
t0\tx269 \up0 \expndtw0\charscalex102 Alamprese, C., Foschino, R., Rossi, M., Po
mpei, C., & Savani, L. (2002). Survival of \line\tab \up0 \expndtw0\charscalex1
02 Lactobacillus johnsonii La1 \up0 \expndtw0\charscalex115 and the influenc
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p0 \expndtw0\charscalex107 manufactured ice-cream produced with different sugar
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\expndtw0\charscalex103 ANVISA (2008). Brazilian Agency of Sanitary Surveillance
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br/alimentos/comissoes/tecno_lista_alega.htm}}}\ul0\nosupersub\cf7\f8\fs13 >. \u
p0 \expndtw0\charscalex102 Arai, O., Sakaki, M., & Sugimoto, T. (1996). Effectiv
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p0 \expndtw-1\charscalex100 Basyg\u728?it, G., Kuleasan, H., & Karahan, A.
G. \up0 \expndtw0\charscalex105 (2006). Viability of human-derived\par\pard
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tw0\charscalex101 Industrial Microbiology and Technology, 33(9), 796-800.\par\pa
rd\ql \li20\sb10\sl-149\slmult0 \up0 \expndtw0\charscalex100 Bolliger, S., Gof
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b \up0 \expndtw0\charscalex100 (Passiflora edulis) and guava (Psidium guajava) p
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rigerated mousses. Brazilian Journal of Microbiology, 38(2), 315-317. \line \up0
\expndtw0\charscalex104 aglar, E., Kuscu, O., Kuwetli, S., Cildir, S. K., Sandal
li, N., & Twetman, S. (2008). \line\tab \up0 \expndtw0\charscalex107 Short-term
effect of ice-cream containing Bifidobacterium lactis Bb-12 on the \line \tab
\up0 \expndtw0\charscalex102 number of salivary mutants streptococci and l
actobacilli. Acta Odontologica \line \tab \up0 \expndtw0\charscalex100 Scandin

avica, 66(3), 154-158.\par\pard\ql \li21\ri646\sb1\sl-159\slmult0\tx269\tx269 \u
p0 \expndtw0\charscalex104 Champagne, C. P., Gardner, N. J., & Roy, D. (2005).
Challenges in the addition of \line\tab \up0 \expndtw0\charscalex102 probiotic c
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tab \up0 \expndtw0\charscalex110 61-84.\par\pard\ql \li21\ri647\sb0\sl-159\slmul
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ristiansen, J. R., & Nielsen, E. W. (1996). Some \line\tab \up0 \expndtw0\chars
calex100 properties of ice cream containing Bifidobacterium bifidum and
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haft, 51, 502-504.\par\pard\ql \li21\ri647\sb0\sl-159\slmult0\tx269\tx269 \up0 \
expndtw0\charscalex103 Collado, M. C., & Sanz, Y. (2005). Induction of acid resi
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7.\par\pard\ql \li21\sb9\sl-149\slmult0 \up0 \expndtw0\charscalex100 Correia, R
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69\tx269 \up0 \expndtw0\charscalex103 Technological and structural aspects. Higi
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\expndtw0\charscalex107 Probiotic culture survival and implications in fermente
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tx269 \up0 \expndtw0\charscalex106 Food and Agriculture Organization of United N
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HO (2001). Evaluation of health and nutritional properties of probiotics \line \
tab \up0 \expndtw0\charscalex111 in food including powder milk with live lactic
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expert consultation, Crdoba, Argentina. <{\field{\*\fldinst {HYPERLINK http
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b\cf15\f16\fs13 ftp://ftp.fao.org/es/esn/}}} \line \tab \up0 \expndtw0\charscale
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port_en.pdf }}{\fldrslt {\ul0\nosupersub\cf15\f16\fs13 food/probioreport_en.pdf}
}}\ul0\nosupersub\cf7\f8\fs13 >.\par\pard\ql \li21\ri646\sb0\sl-159\slmult0\fi0\
tx269\tx269\tx269\tx269\tx269\tx269\tx269 \up0 \expndtw-2\charscalex100 Fvaro-Tri
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07). \line\tab \up0 \expndtw0\charscalex105 Effects of culture, pH and fat
concentration on melting rate and sensory \line \tab \up0 \expndtw0\charsc
alex104 characteristics of probiotic fermented yellow mombin (Spondias mombin L)
ice-\line \tab \up0 \expndtw0\charscalex100 creams. Food Science and Technology
International, 13(4), 285-291. \line \up0 \expndtw0\charscalex100 Fvaro-Trindade
, C. S., Bernadi, S., Boldini, R. B., & Balieiro, J. C. C. (2006). Sensory \line
\tab \up0 \expndtw0\charscalex108 acceptability and stability of probiotic
microorganisms and vitamin C in \line \tab \up0 \expndtw0\charscalex100 ferm
ented acerola ice-cream. Journal of Food Science, 71(6), 492-495. \line \up0 \ex
pndtw0\charscalex104 Gibson, G. R., & Roberfroid, M. B. (1995). Dietary modulati
on of the human colonic \line\tab \up0 \expndtw0\charscalex104 microbiota: Intr
oducing the concept of prebiotics. The Journal of Nutrition, \line \tab
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9\slmult0\tx269\tx269\tx269 \up0 \expndtw0\charscalex105 Gill, C. O. (2006). Mi
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(2003). Viability and survival of free, encapsulated \line\tab \up0 \expndtw0\c
harscalex102 and co-encapsulated probiotic bacteria in ice-cream. Milchwissensch
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\sb9\sl-149\slmult0 \up0 \expndtw-1\charscalex100 Godward, G., Sultana, K., Kail
asapathy, K., Peiris, P., Arumugaswamy, R., & Reynolds,\par\pard\qj \li259\ri647
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x103 bacteria in dairy foods. Milchwissenschaft, 55(8), 441-445.\par\pard\qj \li
20\ri647\sb0\sl-159\slmult0\tx269 \up0 \expndtw0\charscalex103 Goff, H. D. (2008
). 65 years of ice-cream science. International Dairy Journal, 18(7), \line\tab
\up0 \expndtw0\charscalex108 754-758.\par\pard\qj \li20\ri647\sb0\sl-159\slmult0
\tx269 \up0 \expndtw0\charscalex103 Goff, H. D., & Griffiths, M. W. (2006
). Major advances in fresh milk and milk \line\tab \up0 \expndtw0\charsca
lex104 products: Fluid milk and frozen desserts. Journal of Dairy Scien
ce, \up0 \expndtw0\charscalex100 89(4),\par\pard\ql \li258\sb9\sl-149\slmult0 \u
p0 \expndtw0\charscalex107 1163-1173.\par\pard\ql \li20\ri647\sb2\sl-159\slmult0
\tx268\tx269\tx269 \up0 \expndtw0\charscalex108 Haynes, I. N., & Playne, M. J. (
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par\pard\li4112\sb0\sl-149\slmult0\par\pard\li4112\sb0\sl-149\slmult0\par\pard\l
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xpndtw0\charscalex108 International Dairy Foods Association\tab \up0 \expn
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{HYPERLINK http://www.idfa.org/facts/icecream.cfm }}{\fldrslt {\ul0\nosupersub\c
f15\f16\fs13 http://www.idfa.org/facts/}}}\par\pard\ql \li1089\sb10\sl-149\slmul
t0 \up0 \expndtw-1\charscalex100 {\field{\*\fldinst {HYPERLINK http://www.idfa.o
rg/facts/icecream.cfm }}{\fldrslt {\ul0\nosupersub\cf15\f16\fs13 icecream.cfm}}}
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ectd\fs24}

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