all experimental procedures were in accordance with the guidelines of the institutional 
 animal ethical committee .
animal maintenance and handling were in accordance with the indian 
 national institute of nutrition guidelines 
 .
all diets were formulated according to american institute of nutrition ( ain)-93 g 
 standards , which have been recommended for pregnant and lactating female 
 rats 
 .
the composition of the c diet was the same as ain-93 g and contained 
 18  g protein/100  g diet ( table 1 ) .
we formulated 
 two iso - energetic treatment diets low in protein ( 9  g/100  g diet ) by increasing the amount 
 of maize starch to 488  g/100  g diet .
of these diets , one ( low protein ( lp ) ) had soyabean 
 oil ( 7  % ) while the other ( low protein dha  +  ara supplemented ( lps ) ) had a combination of 
 single cell oils containing 4045  % of dha and ara ( martek biosciences ) with soyabean as 
 the basal oil .
the percentages of these single cell oils were calculated based on oral 
 diet safety studies 

 .
special attention was given 
 while processing the lps diet , which was dried at room temperature ( 30c ) to avoid the 
 oxidation of added oils .
the dry pellets were stored at refrigeration temperature ( 4c ) 
 and used within 1 week .
table 1.composition ( g/100 g ) of control and experimental diets ( mean values and standard 
 deviations)diets  clplpsmean 
 sd 
 mean 
 sd 
 mean 
 sd 
 diet composition*carbohydrate603046890568904protein1820294039403fatsoybean oil750274025602arasco00000000081200dhasco0000000000056000vitamin mix100010001000mineral mix||350035003500fatty acid composition ( % ) 16 : 01101117618 : 042412818 : 1n-922222015018 : 2n-654054130618 : 3n-377754220 : 4n-600003522 : 6n-3000024n-6:n-37272509c , control diet ; lp , low - protein diet ; lps , low - protein diet supplemented with 
 dha and arachidonic acid . * proximate analysis ( ten batches of diets per group). arasco , arachidonic acid single cell oil ( martek biosciences). dhasco , docosahexaenoic acid single cell oil ( martek biosciences). mineral mixture ( g / kg mix ) : calcium carbonate 357 , potassium phosphate 196 , 
 potassium citrate 7078 , sodium chloride 78 , potassium sulfate 466 , magnesium 
 oxide 24 , ferric citrate 606 , zinc carbonate 165 , manganous carbonate 063 , 
 cupric carbonate 03 , potassium iodate 001 , sodium selenate 001 , ammonium 
 paramolybdate 0007 , sodium metasilicate 145 , chromium potassium sulfate 0275 , 
 lithium chloride 001 , boric acid 008 , sodium fluoride 006 , nickel carbonate 
 003 , ammonium vandate 0006 , sucrose 22102|| vitamin mixture ( g / kg mix ) : nicotinic acid 3 , calcium pantothenate 16 , 
 pyridoxine - hcl 07 , thiamin - hcl 06 , riboflavin 06 , folic acid 02 , 

d - biotin 002 , vitamin b12 ( 01% in mannitol ) 25 , vitamin 
 e15 , vitamin a 08 , vitamin d 025 , vitamin k 0075 , sucrose 
 974655. fatty acid composition of extracted fat was analysed by gas chromatography , 
 expressed as median values .
composition ( g/100 g ) of control and experimental diets ( mean values and standard 
 deviations ) c , control diet ; lp , low - protein diet ; lps , low - protein diet supplemented with 
 dha and arachidonic acid .
 dhasco , docosahexaenoic acid single cell oil ( martek biosciences ) .  mineral mixture ( g / kg mix ) : calcium carbonate 357 , potassium phosphate 196 , 
 potassium citrate 7078 , sodium chloride 78 , potassium sulfate 466 , magnesium 
 oxide 24 , ferric citrate 606 , zinc carbonate 165 , manganous carbonate 063 , 
 cupric carbonate 03 , potassium iodate 001 , sodium selenate 001 , ammonium 
 paramolybdate 0007 , sodium metasilicate 145 , chromium potassium sulfate 0275 , 
 lithium chloride 001 , boric acid 008 , sodium fluoride 006 , nickel carbonate 
 003 , ammonium vandate 0006 , sucrose 22102 || vitamin mixture ( g / kg mix ) : nicotinic acid 3 , calcium pantothenate 16 , 
 pyridoxine - hcl 07 , thiamin - hcl 06 , riboflavin 06 , folic acid 02 , 
 d - biotin 002 , vitamin b12 ( 01% in mannitol ) 25 , vitamin 
 e15 , vitamin a 08 , vitamin d 025 , vitamin k 0075 , sucrose 
 974655 .
 fatty acid composition of extracted fat was analysed by gas chromatography , 
 expressed as median values .
the animal house at agharkar research institute ( pune , india ) is registered under the 
 committee for the purpose of control and supervision of experiment on animals , india , and 
 follows vigorously all instructions for adequate animal care .
nulliparous female wistar rats weighing 200250  g were obtained from the in - house 
 colony of the institute .
the 
 animals were maintained at 22c on a controlled 12  h light and 12  h dark cycle with an 
 appropriate ventilation system .
rice husk was used as bedding material and was 
 non - nutritive and non - toxic .
when 
 they reached body weights of 200  g they were kept for breeding .
mating was confirmed by 
 sperm - positive vaginal smear . on confirmation of pregnancy , i.e. 0  d of gestation ,
they 
 were randomly allocated to one of the three diet groups ( ten per group ) . in both the c and 
 lp groups ,
the dams 
 received the dietary treatment during gestation ( 21  d ) and lactation ( 21  d ) .
daily food intake during pregnancy and weekly body weights were recorded for dams in all 
 three groups .
dam weights after parturition , 
 litter size and litter weights at birth were recorded .
large litters were reduced to eight 
 pups on day 1 to ensure a standard litter size per dam .
brain fatty acids were analysed for 
 a sub - sample of pups at postnatal day 1 and during lactation ( day 14 ) .
fresh whole brains 
 were collected from anaesthetised pups and cryo - frozen at 80c .
hydrolysis and 
 transesterification of lipids were carried out according to the method of o'fallon 
 et al . 
 .
heptanoic acid ( 17 : 0 ; sigma ) was used as an internal standard .
the 
 methyl esters were extracted with hexane and the fatty acid composition of the extract was 
 analysed by gc using shimadzu model gc-17a version 3 fitted with capillary column spb 35 . 

the analysis conditions were as follows : capillary column , 30  m    032  mm inner diameter , 
 film thickness , 0.25 m .
the temperature programme was as follows : the initial temperature 
 100c ( 5  min hold ) , 100220  c ( 10c per min ) .
gc parameters were : injector temperature 
 200c , detector temperature 220c , flame ionisation detector , carrier gas he ; flow rate 
 18  ml / min ; split ratio 33:1 . for milk collection
, pups were separated from their dams after providing a sufficient 
 time for nursing the young ones . on day 1 of lactation ,
two pups / litter / diet group were 
 randomly selected for collection of gastric milk samples . on day 14 of lactation too , the 
 gastric milk sample from one pup / litter / diet group was randomly selected . before 
 collection of milk
, pups were exposed to anaesthetic ether and then milk was obtained from 
 the stomach and the collected samples were cryo - frozen at 80c .
the milk lipids were 
 extracted and transesterified according to the method of christopherson & 
 glass 
  and analysed by gc using shimadzu model gc-17a version 3 fitted with 
 capillary column spb 35 .
the analysis conditions were as follows : capillary column , 
 30  m    032  mm inner diameter , film thickness , 0.25 m .
the temperature programme was as 
 follows : initial temperature 40c ( 5  min hold ) , 40100c ( 6c / min ) and 100220c 
 ( 10c / min ) .
the carrier gas was he with a flow rate of 
 18  ml / min and a split ratio of 33:1 .
fatty acid concentrations were calculated from their 
 relative peak areas as compared with the internal standard .
individual fatty acids were identified by comparing the 
 retention time of the peak with that of standard fatty acid methyl esters ( 37-component 
 fame mix ; sigma ) .
the treatment groups were compared 
 with the c group by one - way anova and the post hoc bonferroni test was 
 applied to make pair - wise comparisons . the kruskal 
wallis independent sample 
 non - parametric analysis was also applied to test significance in the case of small sample 
 size ( n  <  30 ) , for the comparison of brain and milk fatty acids 
 profiles .
the data were analysed using the spss / pc+ package ( version 11.0 ; spss inc . ) .
despite differences in protein levels , there was no difference in food intake during 
 pregnancy ( table 2 ) and lactation among the dams 
 from the c and treatment groups .
the total weight gain during pregnancy was the highest in 
 the dams from the c group ; however , it did not differ significantly from those in the lp 
 and lps groups ( table 2 ) .
table 2.effect of dietary treatment on food consumption , body weight gain and reproductive 
 performance of control diet - fed ( c ) and experimental dams ( mean values and standard 
 deviations)diet group 
clplpsmean 
 sd 
 mean 
 sd 
 mean 
 sd 

 p 
 no . of dams9910food consumption ( g / week)gestation week 1140525511981761228254nsgestation week 2135532511112101096199nsgestation week 310491541037151931274nspre - pregnancy weight ( g)219315520591722280277nsweight gain ( g)gestation week 1176321502215432nsgestation week 2194461593015528nsgestation week 3150221482113339nstotal gain875185836102819214nsreproductive outcomelitter size10623103369435nslitter weight ( g)564143550156483142nsbirth weight ( g)540555055307nslp , low - protein diet ; lps , low - protein diet supplemented with dha  +  arachidonic 
 acid .
effect of dietary treatment on food consumption , body weight gain and reproductive 
 performance of control diet - fed ( c ) and experimental dams ( mean values and standard 
 deviations ) lp , low - protein diet ; lps , low - protein diet supplemented with dha  +  arachidonic 
 acid .
diet had no effect on litter size 
 and litter weights at birth ( table 2 ) .
the fatty acid composition of pups ' brains on postnatal day 1 and during lactation ( day 
 14 ) is shown in table 3 . at birth
, there was no 
 difference in the level of sfa between groups ; however , the level of mufa was 
 significantly higher in brains of pups from the lp group compared to pups from the c and 
 lps groups . among the essential fatty acids , the level of la was similar between groups 
 but that of ala was significantly higher in pups of the lps group . among the supplemented 
 fatty acids , the level of ara in brain lipids did not differ between groups , but dha was 
 significantly higher in the pups from the lps group .
in addition , pups from this group had 
 a significantly higher amount of epa ( 20 : 5n-3 ) and total 
 n-3 and lower level of n-6 docosapentaenoic acid ( 22 : 
 5 ) as compared with the lp group .
consequently , the 
 n-6:n-3 ratio for brain fatty acids from lps pups was 
 significantly lower than for pups from the c and lp groups .
table 3.brain lipid fatty acids expressed as percentage total fatty acids of neonatal 
 offspring of dams fed the control diet ( c ) and experimental diets ( mean values and 
 standard deviations)diet group 
clplpsmean 
 sd 
 mean 
 sd 
 mean 
 sd 
 postnatal day 1no .
of pups6910male355female345total sfa453494393644949total mufa341433913535733la ( 18 : 2n-6)180610061007ala ( 18 : 3n-3)110605031306gla ( 18 : 3n-6)170811060504ara ( 20 : 4n-6)911787158818epa ( 20 : 5n-3)080306061508dpa ( 22 : 5n-6)0505080400701dha ( 22 : 6n-3)550941106210total n-6131201172110424total n-3740552188916total pufa205211693719335n-6:n-3170324051203postnatal day 14no . of pups667male334female333total sfa368124626838034total mufa348243266432225la ( 18 : 2n-6)160709021607ala ( 18 : 3n-3)07090010041709gla ( 18 : 3n-6)160708031006ara ( 20 : 4n-6)135041181413513epa ( 20 : 5n-3)200808062209dpa ( 22 : 5n-6)040208020202dha ( 22 : 6n-3)840561079509total n-6171201431416324total n-311113691213417total pufa283162121229740n-6:n-3150221061208lp , low - protein diet ; lps , low - protein diet supplemented with dha  +  arachidonic 
 acid ; la , linoleic acid ; ala , -linolenic acid ; gla , -linolenic acid ; ara , 
 arachidonic acid ; dpa , docosapentaenoic acid . mean values within a row with unlike superscript letters were 
 significantly different ( p<005 ; preplanned comparisons 
 using post hoc tests ) . brain lipid fatty acids expressed as percentage total fatty acids of neonatal 
 offspring of dams fed the control diet ( c ) and experimental diets ( mean values and 
 standard deviations ) lp , low - protein diet ; lps , low - protein diet supplemented with dha  +  arachidonic 
 acid ; la , linoleic acid ; ala , -linolenic acid ; gla , -linolenic acid ; ara , 
 arachidonic acid ; dpa , docosapentaenoic acid
mean values within a row with unlike superscript letters were 
 significantly different ( p<005 ; preplanned comparisons 
 using post hoc tests ) .
by mid - lactation ( day 14 ) , the percentage of ara and dha in brain lipids of pups 
 increased significantly in all diet groups ( fig . 

however , compared with pups from the c and lps groups , brain lipids of pups from 
 the lp group not only had significantly higher sfa ( mainly palmitic ; 16 : 0 2973  % and 
 stearic ; 18 : 0 582  % compared with 2618 and 507  % in pups from the c group ) but also 
 had markedly higher levels of docosapentaenoic acid , which is a marker of 
 n-3 fatty acid deficiency .
on the other hand , with regard to essential 
 fatty acids , lps pups continued to accrue higher levels of ala , ara , epa , dha and total 
 n-3 fatty acids as compared with lp pups .
1.effect of dietary treatment on accretion of arachidonic acid ( ara ) ( a ) and dha ( b ) 
 in whole brain phospholipids of offspring during the early postnatal period ( /// , 
 day 1 ;  , day 14 ) .
values are means , with standard errors represented by vertical 
 bars ( six to ten rats per group ) .
* mean value was significantly different from that 
 at day 1 ( p < 005 ; preplanned comparisons ) .
lp , low - protein 
 diet ; lps , low - protein diet supplemented with dha + ara . effect of dietary treatment on accretion of arachidonic acid ( ara ) ( a ) and dha ( b ) 
 in whole brain phospholipids of offspring during the early postnatal period ( /// , 
 day 1 ;  , day 14 ) .
values are means , with standard errors represented by vertical 
 bars ( six to ten rats per group ) .
* mean value was significantly different from that 
 at day 1 ( p < 005 ; preplanned comparisons ) .
lp , low - protein 
 diet ; lps , low - protein diet supplemented with dha + ara .
the fatty acid composition of dams ' milk at the beginning of lactation ( day 1 ) and 
 mid - lactation ( day 14 ) is shown in table 4 .
on 
 day 1 , there was no difference in the levels of sfa , mufa or essential fatty acids , namely 
 la and ala , between groups .
however , the level of scfa and medium - chain fatty acids 
 ( c4-c12 ) was significantly lower while the levels of both supplemented fatty acids ( ara 
 and dha ) were significantly higher , resulting in a higher amount of total 
 n-3 and pufa in the milk samples of dams from the lps group than those 
 from the c and lp groups .
the n-6:n-3 ratio of milk was 
 the highest in the dams from the lp group .
table 4.milk fatty acids expressed as percentage total fatty acids during early and late 
 lactation in dams fed the control diet ( c ) and experimental diets * ( mean values and 
 standard deviations)diet group 
clplpsmean 
 sd 
 mean 
 sd 
 mean 
 sd 
 lactation day 1no . of pups666male333female333total scfa  +  mcfa14346133434124total mufa278262896230329total sfa494464986343635la ( 18 : 2n-6)186241832518923ala ( 18 : 3n-3)190514042007ara ( 20 : 4n-6)180513033407dha ( 22 : 6n-3)050402021303total n-6204261972122422total n-3240616043511total pufa228242132126128n-6:n-39231134417030lactation day 14no . of pups555male333female223total scfa178131903218337total mufa189132012620016total sfa508055261250415la ( 18 : 2n-6)164101550714806ala ( 18 : 3n-3)200512012905ara ( 20 : 4n-6)1304040042310dha ( 22 : 6n-3)0602006011708total n-6177071590617216total n-3260513014612total pufa203051720621825n-6:n-37118124133909lp , low - protein diet ; lps , low - protein diet supplemented with dha  +  arachidonic 
 acid ; mcfa , medium - chain fatty acids ; la , linoleic acid ; ala , -linolenic acid ; 
 ara , arachidonic acid . mean values within a row with unlike superscript letters were 
 significantly different ( p<005 ; preplanned comparisons 
 using post hoc tests ) . * post - nursing gastric milk samples collected from pup stomachs . milk fatty acids expressed as percentage total fatty acids during early and late 
 lactation in dams fed the control diet ( c ) and experimental diets * ( mean values and 
 standard deviations ) lp , low - protein diet ; lps , low - protein diet supplemented with dha  +  arachidonic 
 acid ; mcfa , medium - chain fatty acids ; la , linoleic acid ; ala , -linolenic acid ; 
 ara , arachidonic acid .  
mean values within a row with unlike superscript letters were 
 significantly different ( p<005 ; preplanned comparisons 
 using post hoc tests ) . * post - nursing gastric milk samples collected from pup stomachs . by mid - lactation ( day 14 )
the percentage of scfa and medium - chain fatty acids increased 
 significantly in the milk of dams from the lps group ( 41183 ; p  =  000 ) 
 and nullified the between - group differences observed on day 1 of lactation . among the 
 essential fatty acids , ala ,
ara , dha , total n-3 and pufa content was the 
 highest in the milk of dams from the lps group , but was the lowest in the case of dams 
 from the lp group . within each dietary group ,
a reduction in dha content of milk from day 
 1 to day 14 was observed but was maximum in the case of the lp group ( 02006 ) .
a similar 
 reduction was observed in the n-6:n-3 ratio , for both 
 the c and lps groups , but more sharply in the case of the latter while it remained 
 constant in the lp group .
correlations between dams ' milk and pup brain fatty acids were computed for day 14 of 
 lactation , by pooling the data for all diet groups ( table 5 ) .
we observed significant positive associations for the 
 n-3 essential fatty acids , i.e. ala but not for the n-6 
 essential fatty acids , namely la .
further , a strong direct association was also noted for 
 the respective n-3 lc - pufa , i.e. dha , but not for the 
 n-6 lc - pufa , i.e. ara .
however , the total n-3 and 
 n-6 levels in milk and brain lipids showed significant positive 
 associations as did the total pufa content .
one interesting observation was the 
 significant inverse association between total sfa in maternal milk and total 
 n-3 content of pup brains ( r 0612 ; 
 p  <  005 ) . table 5.pearson correlation coefficients for dam milk and offspring brain fatty acids 
 during mid - lactation ( day 14)fatty acidcorrelation coefficient 
 ( r)total sfa0.435total mufa0.021la ( 18 : 2n-6)0.133ala ( 18 : 3n-3)0.728**ara ( 20 : 4n-6)0.462*dha ( 22 : 6n-3)0.692**total n-60.595*total n-30.773**total pufa0.755**n-6:n-30.724**la , linoleic acid ; ala , -linolenic acid ; ara , arachidonic acid.significant one - tailed correlation : * p  <  005 , * * 
 p  <  001. pooled sample for all diet groups : n 15 .
pearson correlation coefficients for dam milk and offspring brain fatty acids 
 during mid - lactation ( day 14 ) la , linoleic acid ; ala , -linolenic acid ; ara , arachidonic acid .
significant one - tailed correlation : * p  <  005 , * * 
 p  <  001 .  pooled sample for all diet groups : n 15 .
maternal low protein intake during pregnancy and lactation has been shown to considerably 
 perturb the normal metabolism of essential fatty acids , namely la and ala , resulting in poor 
 ara and dha accumulation in the brain of growing offspring 

 . yet
, there is paucity of studies investigating the effects of 
 supplementation of lc - pufa at low protein levels on changes in brain fatty acid content of 
 the offspring and none have examined the consequent alterations in maternal milk profile .
we 
 observed that in conditions of protein restriction , supplemental dha and ara fed to dams led 
 to their higher incorporation into pup brain lipids by altering the fatty acid profile of 
 dams ' milk . before discussing the important findings of the study it may be worthwhile to mention some 
 of the points .
firstly , although the number of dams allocated to different diet groups was 
 small , it was similar to those reported in earlier studies 



 .
secondly , the number of pups dissected at day 0 and day 14 was modest 
 as it was necessary to maintain a sizeable number of pups / group till adulthood , being a 
 long - term study . nevertheless , non - parametric tests were used for comparing differences in 
 fatty acid profiles .
we observed that even at the low protein level of 9  % , with or without supplementation , 
 litter size or litter weight did not differ significantly from that of the c group .
our 
 observations are in agreement with reports from earlier studies that used diets containing 
 menhaden fish oil 
  or used a large range of la / ala ratios ( 1071035 ) and did not find 
 differences in length of gestation , litter size or pup weights between c and treatment 
 groups 
 .
even a protein - restricted diet with supplemental fish oil fed to 
 pregnant dams did not show adverse effects on gestation , litter size or pup 
 weight 
 .
this has been attributed to the fact that in rats , a considerable part 
 of growth and development occurs after birth than in utero , making them 
 less susceptible to unfavourable nutrient situations while in the womb 
 .
the rate of accumulation of dha and ara in the rat brain is the highest during early 
 postnatal life and supply of these fatty acids during brain growth can boost the rate of 
 brain mass expansion 
 .
we observed a significant increment in total brain ara and dha content 
 from birth to 14  d of postnatal life in pups from all dietary groups ; however , the highest 
 accretion was noted in pups of lps dams .
further , we also noted a strong positive 
 correlation between maternal milk ala , dha , total n-3 content and brain 
 n-3 fatty acid profiles , corroborating the hypothesis that maternal milk 
 fatty acid content indeed determines pup brain fatty acid composition .
two important and independent changes observed in milk fatty acid composition of dams from 
 the lp and lps groups are in need of attention .
firstly , in the case of dams from the lp 
 group , despite comparable levels of n-6 la in their milk , the milk was 
 deficient in its long - chain metabolite , namely ara as compared with its proportion in milk 
 from dams in the c group .
as mentioned earlier , it has been shown that mammary glands 
 actively participate in lc - pufa synthesis during lactation 
 , and so the protein - restricted diet may have down - regulated enzymes 
 involved in desaturation and the elongation pathway of n-6 la causing a 
 reduction in the synthesis of ara .
have reported similar effects of feeding 10  % protein diets to pregnant 
 and lactating rats on the metabolism of essential fatty acids in mammary glands .
the second notable observation was in the case of the lps group which showed a 
 significantly low proportion of scfa and medium - chain fatty acids in dams ' milk during 
 initiation ( day 1 ) of lactation .
however , by mid - lactation ( day 14 ) a significant increase in these 
 fatty acids made the levels comparable with those of the c group , indicating metabolic 
 adaptations . for example , even in human subjects it has been shown that milk fatty acid 
 composition undergoes changes during the course of lactation 
 , which may be a feedback mechanism in order to meet the requirements for 
 postnatal growth of the offspring .
our observation therefore suggests that examining 
 alterations in milk fatty acid composition in the light of pup growth during lactation would 
 be of immense importance .
provision of preformed dha and ara to the pups through milk is crucial for optimal 
 accretion of lc - pufa in the rapidly growing brain of pups , as they are incapable of 
 synthesising them from precursors 
 .
we observed that supplementation of preformed lc - pufa to 
 protein - restricted dams resulted in higher dha and total n-3 fatty acids in 
 their milk , unlike their non - supplemented lp counterparts .
this observation thus calls 
 attention to the significance of lc - pufa supplementation at low protein levels in order to 
 overcome the resulting deficiency due to poor inter - conversion of ala to epa and dha and 
 also to optimise the n-6:n-3 ratio of milk . providing 
 lc - pufa during
protein deficiency also served to conserve maternal ala pools , possibly due 
 to a lower amount of ala entering the biosynthetic pathway , and enhanced epa levels in milk 
 by retro - conversion of dha .
this brought about a significant change in the total 
 n-3 pools and n-6:n-3 ratio of lps milk 
 and led to superior accretion of dha in the brains of lactating pups .
a large body of work suggests that an adequate supply of dha during early life is essential 
 for optimal growth and brain development .
the present findings clearly demonstrate that low 
 maternal dietary protein levels can profoundly affect dha accretion in developing offspring 
 due to n-3-deficient milk , and supplementing dha and ara in such a scenario 
 can positively alter both maternal milk and pup brain fatty acid composition .
the effect of 
 the observed biochemical change on the functional capacity of the brain in the offspring 
 needs to be evaluated .
however , evidence from several supplementation studies in situations 
 of adequate protein suggests that learning and cognitive behaviour are augmented with 
 improvement in brain dha status 


 .
therefore , our findings in a protein - deficient condition assume greater 
 importance owing to the fact that the effects of supplementation are more pronounced in 
 conditions of deficiency .
they emphasise the need for supplying adequate food sources of dha 
 to mothers during pregnancy and lactation to offset the negative effects of maternal protein 
 deficiency on accretion of fetal brain dha and subsequently brain growth .
 long - chain pufa ( lc - pufa ) are important for fetal and neonatal brain development . 
 
 however , their accretion in the brain is compromised during maternal protein restriction . 
 hence , we investigated the effect of maternal supplementation with n-3 
 dha plus n-6 arachidonic acid ( ara ) at a low protein level ( 9  % ) on 
 offspring brain fatty acid accretion using wistar rats ( nine rats per group ) randomly fed 
 a control ( c ) , a low - protein ( lp ) or a low - protein dha  +  ara - supplemented ( lps ) diet 
 during gestation and lactation . at birth , pups from the lps group had the highest brain 
 dha and n-3 fatty acid levels ( p  =  0001 ) , whereas pups 
 from the lp group had the highest mufa ( p  =  005 ) but the lowest dha and 
 total n-3 pufa levels ( p  =  0000 ) . during lactation , 
 pups from the lps group accrued significantly more -linolenic acid 
 ( p  =  0003 ) , epa ( p  =  002 ) and dha 
 ( p  =  0000 ) in brain lipids than pups from the lp group , whereas brain 
 lipids of pups from the lp group had markedly increased levels of the n-3 
 deficiency marker docosapentaenoic acid and n-6:n-3 
 ratio ( p  =  0000 ) . owing to supplementation , milk from lps dams had the 
 highest dha and ara , but lower scfa and medium - chain fatty acids as compared with milk 
 from c and lp dams during early lactation , but normalised by mid - lactation . to conclude , 
 
 adverse effects of restricted maternal protein intake on lc - pufa accretion in the brain of 
 offspring were ameliorated by alterations in maternal milk fatty acid profile due to 
 supplementation . 
 results underscore the importance of lc - pufa for protein - deficient 
 mothers during gestation as well as lactation to achieve the optimum brain lc - pufa status 
 of progeny . 
Materials and methods
Results
Discussion