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Some Effects of Feeding Various Filled Milks to Dairy Calves. II. FecaL Characteristics and Digestibility Data1
SOME EFFECTS OF FEEDING VARIOUS FILLED MILKS DAIRY CALVES. II. FECAL CHARACTERISTICS AND DIGESTIBILITY DATA 1
TO
R. S. ADAMS," J. E. GANDE]~,~ ~1?.W. flUI~]~ICXSON, ~ .~ND J. H. SAUTTEI~ Department of Dairy Husbandry and Veterinary Pathology Section, University of Minnesota, St. Paul SUMMARY Young' dairy calves were fed- low-fat rations or filled milks containing various fats. One group received a corn oil filled milk which was prepared daily rather than weekly, to study effects of storage. All other diets were prepared once each week. Differences in the color of feces were observed among the various rations. Abnormally large amounts of feces were voided by animals fed the corn oil filled milk prepared weekly. Daily p r e p a r a t i o n of the corn oil filled milk diet or the feeding of hydrogenated corn oil greatly reduced the amounts of feces excreted. A p p a r e n t dry matter digestibility for the weekly p r e p a r e d eorn oil filled milk group was significantly less than that obtained for any of the other diets. The a p p a r e n t metabolic fecal fat excretion by calves fed a skimmilk diet containing 0.1% butterfat amounted to 5.36% of the fecal dry matter. I t was established that the a p p a r e n t digestion coefficients for butter oil were significantly greater than those for corn oil, hydrogenated corn oil, or lard. Daily preparation of the corn oil filled milk significantly increased dry matter digestibility, but did not significantly increase f a t digestibility over the weekly p:reparation of this ration. Partition of the fecal fat revealed that a considcrt~blc portion was excreted as soaps by animals receiving the various diets. The fecal fat partition of ealves fed butter oil filled milk was similar to that of apparent mett~bolic fecal fat, as excreted by animals in the skimmilk group. T h e effects of v a r i o u s filh,d m i l k s u p o n the p h y s i e a l c o n d i t i o n , a p p e a r a n e e , a n d g r o w t h of calv~,s have beeu p r e s e n t e d in an e a r l i e r p a p e r ( 1 ) . T h e r e s e a r c h r e p o r t e d h e r e r e p r e s e n t s a f u r t h e r a t t e m p t to a s e e r t a i n some of t h e f a c t o r s r e s p o n s i b l e for t h e u n s a t i s f a c t o r y w e i g h t g a i n s a n d p o o r f e e d u t i l i z a t i o n enc o u n t e r e d when h i g h l y u n s a t u r a t e d vegetatfle oils ~re f e d to d a i r y ealves. F~PER1MI?INTAL PI~()CEDUP~E A n m n b e r of calves f r o m s e v e r a l of the g r o u p s m e n t i o n e d in an e a r l i e r p a p e r were u s e d in t h e d i g e s t i b i l i t y s t u d i e s r e p o r t e d here. D e t a i l s on t h e f e e d i n g a n d m a n a g e m e n t of t h e s u b j e c t s w e r e p r e s e n t e d in a p r e v i o u s r e p o r t ( 1 ) . S i n c e t h e a n i m a l s f r o m s e v e r a l of t h e g r o u p s were h i g h l y s u s c e p t i b l e to r e s p i r a t o r y d i s o r d e r s a ~ d o'ften in a w e a k e n e d c o n d i t i o n , t h e c u s t o m a r y collection p e r i o d u s e d in d i g e s t i o n t r i a l s was n o t e m p l o y e d . I n s t e a d , t h e c a l v e s w e r e p e r i o d i c a l l y p l a c e d ill m e t a b o l i s m s t a l l s for a t h r e e - d a y p e r i o d at a p p r o x i m a t e l y 2 0 - d a y i n t e r v a l s , b e g i n n i n g at :t5 clays of age. T o t a l collection of the feces v o i d e d b y the s u b j e c t s was m a d e d u r i n g the 48 hr. f o l l o w i n g a p r e l i m i n a r y Received for public,qtlon duly 28, 1958. a Scientific Jourmd Series P~per No. 4119, Minnesota. Agrieult.ural Experiment Station. ~Present address: ])'dry Science Extension Section, The Pem~sylvania St~tc University, University Park, Pem~syh, ania. ~Present address: Dep~Jrtment of Agricultural l~ioehemistry, The Ohio State University, Columbus, Ohio. 4 Retired. 1562
EFFECTS OF FEEDING PILLED MILKS TO CALVES. II.
1563
period of 24 hr. Feed intake was held constant for a p p r o x i m a t e l y 1 wk. p r i o r to the collection period. This m a n n e r of collection did not u n d u l y jeopardize the health of the individuals and p r e v e n t e d interference with other phases of the experiment, p a r t i c u l a r l y those dealing with the development of muscular involvement. I t also afforded an o p p o r t u n i t y to determine whether digestibility was progressively affected d u r i n g the experimental period. Following the weighing of the feces voided d u r i n g the collection period, representative samples were taken and stored at --20 ° C. until analyses were performed. The storage period for the fecal samples did not exceed 8 wk. D r y m a t t e r was determined by oven drying. A sample of the dried feces was pulverized and used in the determination of total f a t content and the various fat components. A slight modification in the method of Van der K a m m e r and associates (12) made it possible to ascertain the grams of soaps as well as free f a t t y acids and neutral fat present per 100 g. of fecal d r y matter. The original method gives values for neutral f a t and the total f a t t y acids present as fecal soaps and free f a t t y acids. In the modification an additional titration was p e r f o r m e d on a portion of the dried fecal sample, which was refluxed without the addition of ItC1 to avoid the liberation of the f a t t y acids present as soaps. The value obtained f r o m this additionM titration was taken as the original free f a t t y acids present in the fecal sample. The difference between this vahie and the one obtained after refluxing with acid was taken as the original soaps present in the seraph t of dried feces. I~ESULTS AND DISCUSSION
Fecal characteristics. As mentioned in the earlier report (1), m a r k e d differences in the eonsisten(,y of the feces were f o u n d amoJ~g the various d i e t a r y rogimes. The mean daily fecal excretion values in '['able 1 provide a reasonable TA BLE 1 Fecal o u t p u t by calves on the v'~rious diets
Group l II ]II IV V VI Villi VIII
IX
Diet Corn oil filled nfilk ( p r e p a r e d weekly) Corn oil filled milk (prep:trod daily) l l y d r o g e n a t e d corn oil filled milk L a r d filled milk B u t t e r oil filled milk Fortified skimmilk Fortitied skimmilk plus corn oil Fortified skimmilk plus h y d r o g e n a t e d soybean and cottonseed oil blend Skimmilk
Wet basis.
No. calves
No. tri'ds
Me.m dry matter intake
M e a n daily fecal excretion"
Standard error of m e a n
I0
41
(g.) 829.8
(.q.) 1,170.3
-4- 22.8
4
16
839.1
477.2
-4- 15.1
3 4 3 '2
12 10 12 6
1,161.2 ],256.6 896.8 2,031.8
698.3 557.6 469.2 1,772.8
-4- 23.9 -4-20.6 ~__ 20.6 +--319.7
5
22
1,689.1
],398.1
+
2 2
8 11
2,063.5 667.0
1,428.4 907.8
-4-162.3 +- 55.9
69.0
1564
R . S . ADAMS ET AL
indication of the amount of d i a r r h e a present a m o n g the various groups. The fecal output of subjects in groups with feces of a p p a r e n t l y n o r m a l consistency (Groups I I - V , inclusive) was somewhat greater t h a n t h a t r e p o r t e d b y B l a x t e r and Wood (4) in their s t u d y of diarrhea in calves. Animals fed corn oil filled milk p r e p a r e d weekly were afflicted with a severe diarrhea. I n some trials, as much as 3,000 g. of feces were collected daily f r o m individuals in this group. Tocopherol supplementation of the corn oil filled milk diets did not reduce the severity of the diarrhea. Calves receiving a corn oil filled milk p r e p a r e d daily in an a t t e m p t to reduce its toxicity voided a considerably smaller a m o u n t of feces. H y d r o g e n a t i o n of the corn oil g r e a t l y reduced the amount of feces excreted daily. Similarly to Group I, the feces of animals on the fortified skimmilk plus corn oil diet were extremely loose. Although the mean daily excretion of feces was greatest for the fortified skimmilk group, there was a large amount of variation in the values for t h a t group, as shown b y the s t a n d a r d error of the mean. Considerable v a r i a t i o n in the consistency of the feces was also noted in the trials of animals in Group I X . The m a r k e d fecal excretion b y calves fed the various fortified skimmilk diets undoubtedly was influenced to some extent by the a b n o r m a l l y high solids content of these rations. The factors responsible for the relatively large fecal output b y the skimmilk group are not a p p a r e n t . I t is interesting to note, however, t h a t I o w a workers (13) recently r e p o r t e d that diets high in concentrations of milk solids-not-fat a n d / o r low in f a t are likely to cause diarrhea. Differences in the color of feces were also quite pronounced a m o n g the variot~s experimental rations. The feces of animals on the weekly p r e p a r e d corn oil filled milk diet were characteristically greenish-yellow in color; whereas, those excreted by calves receiving the same diet p r e p a r e d daily were a darker green. The excretions of the lard filled milk group were usually grayish-blue or sometimes clay-like. Subjects on the whole milk, skimmilk, fortified skimmilk, a n d b u t t e r oil filled milk rations voided feces which were the usual brown in color. I n contrast, the fecal excretions of individuals on the h y d r o g e n a t e d vegetable oil filled milks were invariably gray. Differences in bile secretion, intestinal f a t excretion, intestinal flora, and the oxidation-reduction potential of the digestive tract, as well as other factors, m a y influence color of feces.
Digestibility data. The a p p a r e n t d r y m a t t e r digestion coefficients for several of the v,~rious diets may be f o u n d in Table 2. These data were t r e a t e d statistically TABLE 2 A p p a r e n t d r y m;~tter d i g e s t i o n coefficients of t h e v a r i o u s e x p e r i m e n t a l diets
NO.
2~'o.
Diet
c~lves
trials
Corn oil filled milk prepared weekly Corn oil filled milk prepared daily t[ydrogenated corn oil filled milk Lard filled milk Butter oil filled milk
8 4 3 4 3
41
16 12 10 12
Av. dry matter digestion coefficient 81.1 90.4: 89.5 90.9 93.6
EFFECTS
OF FEEDING
FILLED
MILKS
TO CALVES.
II.
1565
according to methods outlined b y Snedecor (11) and D u n c a n (9, 10), to test the differences a m o n g the various t r e a t m e n t means at the 5% level of significance. The average d r y m a t t e r digestion coefficient for the weekly p r e p a r e d corn oil filled milk group was found to be significantly less t h a n t h a t f o r a n y one of the other diets. No other significant differences in a p p a r e n t d r y m a t t e r digestibility were noted among the various diets. The d r y m a t t e r digestion coefficients for calves in the various fortified skimmilk groups were not included in the analyses because of the limited amount of data available for these diets. The significantly greater a p p a r e n t d r y m a t t e r digestibility of the daily prep a r e d corn oil filled milk ration is of considerable interest. Undoubtedly, a large portion of the increase in a p p a r e n t d r y m a t t e r digestibility m a y be associated with the decrease in fecal excretion and degree of diarrhea. As previously noted, the feces of calves in the daily p r e p a r e d corn oil filled milk group were n e a r l y normal in consistency. Blaxter and Wood (4) reported t h a t the a p p a r ent digestibility of fat, protein, and ash drops f r o m as high as 97 to as low as 40% in some instances, as the result of diarrhea in calves. The a p p a r e n t f a t digestibility data are presented in Table 3. The adjusted fat digestion coefficients listed in this table are values adjusted for metabolic fecal fat as follows: The mean a m o u n t of f a t excreted daily by calves in the skimmilk group was expressed as a percentage of the fecal d r y matter. Since the diet of these aninm]s contained only 0.1% fat, it was a ~ u m e d t h a t this value, 5.36%, was indicative of the metabolic fecal fat expressed as a percentage of the fecal d r y matter. Thus, an amount equivalent to 5.36% of the fecal d r y m a t t e r was deducted from the total fat excretion in calculating the adjusted f a t digestion coefficient for calves receiving the sew~,ral filled milks. The data used in determining the metabolic fecal fat excretion were derived f r o m 11 digestion trials conducted with two subjects in the skimmilk group. The finding that the metabolic fecal f a t excretion by calves in this group a m o u n t e d to 5.36% of the fecal d r y m a t t e r is in v e r y close agreement with the 5.30% value obtained under similar conditions by previous workers (5). More recently, C u n n i n g h a m and Loosli (7) re,ported values of 3 . l - 5 . 8 % when the metabolic fecal fat excreted b y calves on a fat-free purified diet was expressed as a percentage of the fecal d r y matter. A?l analysis of variance of the u n a d j u s t e d fat digestibility data indicated that significant diffcre, nces existed among the various diets. F u r t h e r statistical t r e a t m e n t revealed that the digestibility of b u t t e r oil was superior to that of tile fats in each of the other diets studied. I n every ease this superiority of b u t t e r oil was significant at the 1% level. No other significant differences in unadjusted fat digestion coefficients existed among the remaining diets. Signifieant differences were f o u n d to exist also among the adjusted fat digestion coefficients for tile various rations. The digestibility of butter oil was again demonstrated to be significantly greater (1% confidence level) t h a n that of the fats in the other diets. No other significant differences were noted a m o n g the remaining groups. ]it shouht be pointed out that both the highest and lowest f a t digestion coefficients for the over-all experiment occurred in the weekly
TABLE Summary
Diet
of apl)arent
No. calves
fat digestibility
No. trials
3
data for calves on the various experimental
Av. unadjusted fat digestion coefficient
Av. adjusted " fat digestion coefficient
diets
Free fatty aelds
Soaps
Neutral fat
(av. % fecal fat) C o r n oil filled milk prepared weekly C o r n oil filled m i l k prepared daily Hydrogenated c o r n oil filled m i l k L a r d filled m i l k B a t t e r oil filled m i l k Skimmilk
8
32
84.3
88.3 +__ 0.304
22.9 _ 0.53 b
4 2 . 8 ~ 0.85 b
34.3 ___ 0 . 4 8 b
4
14
88.5
90.2 _4- 0.36
1 2 . 7 __+ 0.87
57.7 + 1.20
2 9 . 6 __+ 0.73
3 4 3 2
10 7 9 8
88.2 S7.1 95.1 --
90.3 _+ 0 . 5 4 88.9 ~ 1.06 96.8 _+ 0.26 --
25.7 20.2 25.3 24.2
53.7 52.6 34.3 26.6
20.6 27.2 40.4 49.2
" Values adjusted for metabolic fecal fat as noted in text. ~' M e a n _+ s t a n d a r d e r r o r o f t h e m e a n .
_ 2.26 _ 2.85 ___ 2.62 _ 3.23
~__ 2.22 ~__ 2.99 _.+ 1.70 ~ 2.92
+__ 1.23 -4- 0.79 ___ 1 . 6 9 ___ 2 . 5 7
EFFI~CT8 OF FI~;FDING ~ ] L L E D
M I L K S TO CALVES.
1I.
1567
p r e p a r e d corn oil filled milk group. No consistent effect of collection period upon fat or d r y matter digestibility was observed in any of the groups. I t is interesting to note that the apparent digestibility of the hydrogenated corn oil did not differ significantly from that of corn oil or lard. The melting point range of the hydrogenated corn oil used in this investigation was 4 5 48 ° C. Studies reported by Crockett and Dcuel (6), as well as by others, have indicated that the critical melting point range above which there appears to be a marked decrease in fat digestibility is 50-55 ° C. The a p p a r e n t digestion coefficients for hydrogenated corn oil obtained in this experiment compare favorably with those obtained by Augur, Rollman, and Deuel (2), when rats were fed hydrogenated cottonseed oil with a similar melting point. They are, however, somewhat greater than those for hydrogenated soybean oil as reported for calves by de Man (8). Tile digestibility of lard in this study is somewhat less than that reported by Crockett and Deuel (6) for the rat, but somewhat greater than the 82.8% figure obtained by Cunningham and Loosli (7), when calves were fed a purified diet. The adjusted digestion coefficient of 96.8 for b u t t e r f a t is somewhat greater than the 93.0 figure for calves obtained similarly by earlier workers (5). However, both the adjusted and unadjusted digestion coefficients for b u t t e r f a t are in good agreement with values reported by de Man (8) as well as by Blaxter and Wood (3). It should be pointed out that the daily preparation of the corn oil filled milk did not significantly increase the digestibility of this highly unsaturated vegetable oil, although d r y matter dig~stib.ility was sigmifieantly improved, as noted earlier. The partition of fecal fats is also' presented in Table 3. The finding that a considerable portion of the fecal fat is present as soaps is in agreement with previous results obtained with rats (2, 6) and calves (3). Thus, it appears important that fat determinations be made on acidified samples of fecal d r y matter in fat digestibility studies, in order to include fecal soaps in the total fat excretion. It should be noted that a larger portion of fecal fat was excreted as soaps than as neutral fats and free f a t t y acids combined, by animals on the hydrogenated corn oil diet. Angur, Rollman, and l)euel (2) also showed this to be true in digestibility studies of hydrogenated cottonseed oil with rats. This same pattern of h~;al fat excretion occurred in the daily p r e p a r e d corn oil and lard filb,d milk groups, and was closely approximated b y animals on tim weekly prepared corn oil filled milk ration. Blaxter and Wood (4) obtained similar results with calves fed margarin('. A marked d~,~'rease in the percentage of fecal fat excreted as free f a t t y acids occurred when the corn oil filled milk diet was prepared daily. Undoubtedly, a portion of this decrease is associated with the abatement of the severe diarrhea in this group. Blaxter and Wood (4) demonstrated that a large increase in f a t t y acid excretion results in ~,alves afflicted with diarrhea. Regarding the partition of the metabolic fecal fat as listed under the skimmilk diet in Table 3, the percentage of fecal fat excreted as free f a t t y acids closely agrees with the value obtained by Cannon, Espe, and Waide (5). The percentage of metabolic :fecal fat present as neutral fat, however, is approximately 10% greater than in
1568
~. s. ADAMS ET AL
the results reported by these workers. On the other hand, the portion of metabolic fecal fat present as neutral fat in this investigation is in agreement with the results obtained by Cunningham and Loosli (7). It is interesting to n o t e t h e s i m i l a r i t y b e t w e e n t h e f e c a l f a t p a r t i t i o n o n t h e b u t t e r oil f i l l e d m i l k diet and that of what has been assumed to be metabolic fecal fat, as excreted by subjects in the skimmilk group. ACKNOWLEDGMENTS This research was supported in p a r t by a g r a n t from the National Dairy Council on behalf of the American Dairy Association. The authors wist~ to express their appreciation to D. E. Jones and R. L. Northrup for technical assistance. REFERENCES (1) ADAMS, R. S., GULLICKSON, T. W., GANDER, ft. E., AND SAUTTEI~, J. H. Some Effects of Feeding Various Filled Milks to Dairy Calves. I. Physical Condition and Weight Gains, with Special Reference to Low-Fat Rations. J. Dairy Sc~., 42: 1552. 1959. (2) AUGER, V., ROLLMAN, H. S., AND DEUEL, H. H., JR. The Effect of Crude Lecithin on the Coefficient of Digestibility and :Rate of Absorption of Fat. J. Nutrition, 33: 177. 1947. (3) BLAXTER, 1(. L., ).Nl) WOOD, W. A. The Nutrition of the Young Ayrshire Calf. 5. The Nutritive Value of Cow's Whole Milk. Brit. J. Nutrition, 5: 1. 1952. (4) ]~JLAXTER, ]((. L., AND WO01), W. A. Sonic O1)servations on the Biochemical and Physiological Events Associated with ])iarrima in Calves. Vet. Record, 65: 896. 1953. (5) CANNON, C. Y., ESPE, 2). L., AND WA]DE, J. B., JR. The Relation of Dietary F a t and F a t Derivatives in the Feces of Young Dairy Calves. Proc. Am. Soc. Animal Prod~lction, 1932: 85. ]933. (6) CI~OCKETT, M. E., AND DEUI.~L, ]-[. J., JR. A Comparison of the Coefficient of Digestibility and the ]late of Absorption of Several Natur'Jl and Artificial Fats as Influenced by Melting Point. J. Nutrition, 33: ]87. ]947. (7) CUNNIN(]IIAM, H. M., AND LOOSLI, J. K. The Effect of F a t Free Diets on Young Dairy Calves with Observations on Metabolic Fecal F a t and Digestion Coefficients for Imrd •rod Hydrogenated Coconut Oil. J. Dairy S(:i., 37: 453. 1954. (8) ])EMAN, T~I. J. Kalveropfokproef Met G(q~ydeerde Soya-Olic. Tijdschr. l)i~rgeneesk, 75: 175. 1951. (9) DUNCAN, D. ]~. A Significance Test for Differences Y~etween Ranked Treatments in an Analysis of Variance. Virginia J. Sci., 2: 171. ].(}51. (10) DUNCAN, D. B. On the Properties of the Multiple Comparisons Test. Virginia J. Sci., 3 : 49. 1952. (11) SNEr)~;eO~, G. W. Statistical Methods. 4th ed. The Iowa State College Press, Ames, Iowa. 1946. (12) VAN l)ng KAMMER, d. ~[[., HUININK, H. TEN ]~., AND WEYERS, H. A. R~pid Mc~itod for Determining F a t in Feces. J. Biol. Chew., 177: 347. 1949. (13) WING, J. M., JA(;OmUON, N. L., AN~ AH,EN, l~. S. The Effect of Various Restricted Diets on the Growth and on Certain Blood Components of Young ])airy Calves. J. Dairy Sci., 38: 1006. 1.(}55.
TO
R. S. ADAMS," J. E. GANDE]~,~ ~1?.W. flUI~]~ICXSON, ~ .~ND J. H. SAUTTEI~ Department of Dairy Husbandry and Veterinary Pathology Section, University of Minnesota, St. Paul SUMMARY Young' dairy calves were fed- low-fat rations or filled milks containing various fats. One group received a corn oil filled milk which was prepared daily rather than weekly, to study effects of storage. All other diets were prepared once each week. Differences in the color of feces were observed among the various rations. Abnormally large amounts of feces were voided by animals fed the corn oil filled milk prepared weekly. Daily p r e p a r a t i o n of the corn oil filled milk diet or the feeding of hydrogenated corn oil greatly reduced the amounts of feces excreted. A p p a r e n t dry matter digestibility for the weekly p r e p a r e d eorn oil filled milk group was significantly less than that obtained for any of the other diets. The a p p a r e n t metabolic fecal fat excretion by calves fed a skimmilk diet containing 0.1% butterfat amounted to 5.36% of the fecal dry matter. I t was established that the a p p a r e n t digestion coefficients for butter oil were significantly greater than those for corn oil, hydrogenated corn oil, or lard. Daily preparation of the corn oil filled milk significantly increased dry matter digestibility, but did not significantly increase f a t digestibility over the weekly p:reparation of this ration. Partition of the fecal fat revealed that a considcrt~blc portion was excreted as soaps by animals receiving the various diets. The fecal fat partition of ealves fed butter oil filled milk was similar to that of apparent mett~bolic fecal fat, as excreted by animals in the skimmilk group. T h e effects of v a r i o u s filh,d m i l k s u p o n the p h y s i e a l c o n d i t i o n , a p p e a r a n e e , a n d g r o w t h of calv~,s have beeu p r e s e n t e d in an e a r l i e r p a p e r ( 1 ) . T h e r e s e a r c h r e p o r t e d h e r e r e p r e s e n t s a f u r t h e r a t t e m p t to a s e e r t a i n some of t h e f a c t o r s r e s p o n s i b l e for t h e u n s a t i s f a c t o r y w e i g h t g a i n s a n d p o o r f e e d u t i l i z a t i o n enc o u n t e r e d when h i g h l y u n s a t u r a t e d vegetatfle oils ~re f e d to d a i r y ealves. F~PER1MI?INTAL PI~()CEDUP~E A n m n b e r of calves f r o m s e v e r a l of the g r o u p s m e n t i o n e d in an e a r l i e r p a p e r were u s e d in t h e d i g e s t i b i l i t y s t u d i e s r e p o r t e d here. D e t a i l s on t h e f e e d i n g a n d m a n a g e m e n t of t h e s u b j e c t s w e r e p r e s e n t e d in a p r e v i o u s r e p o r t ( 1 ) . S i n c e t h e a n i m a l s f r o m s e v e r a l of t h e g r o u p s were h i g h l y s u s c e p t i b l e to r e s p i r a t o r y d i s o r d e r s a ~ d o'ften in a w e a k e n e d c o n d i t i o n , t h e c u s t o m a r y collection p e r i o d u s e d in d i g e s t i o n t r i a l s was n o t e m p l o y e d . I n s t e a d , t h e c a l v e s w e r e p e r i o d i c a l l y p l a c e d ill m e t a b o l i s m s t a l l s for a t h r e e - d a y p e r i o d at a p p r o x i m a t e l y 2 0 - d a y i n t e r v a l s , b e g i n n i n g at :t5 clays of age. T o t a l collection of the feces v o i d e d b y the s u b j e c t s was m a d e d u r i n g the 48 hr. f o l l o w i n g a p r e l i m i n a r y Received for public,qtlon duly 28, 1958. a Scientific Jourmd Series P~per No. 4119, Minnesota. Agrieult.ural Experiment Station. ~Present address: ])'dry Science Extension Section, The Pem~sylvania St~tc University, University Park, Pem~syh, ania. ~Present address: Dep~Jrtment of Agricultural l~ioehemistry, The Ohio State University, Columbus, Ohio. 4 Retired. 1562
EFFECTS OF FEEDING PILLED MILKS TO CALVES. II.
1563
period of 24 hr. Feed intake was held constant for a p p r o x i m a t e l y 1 wk. p r i o r to the collection period. This m a n n e r of collection did not u n d u l y jeopardize the health of the individuals and p r e v e n t e d interference with other phases of the experiment, p a r t i c u l a r l y those dealing with the development of muscular involvement. I t also afforded an o p p o r t u n i t y to determine whether digestibility was progressively affected d u r i n g the experimental period. Following the weighing of the feces voided d u r i n g the collection period, representative samples were taken and stored at --20 ° C. until analyses were performed. The storage period for the fecal samples did not exceed 8 wk. D r y m a t t e r was determined by oven drying. A sample of the dried feces was pulverized and used in the determination of total f a t content and the various fat components. A slight modification in the method of Van der K a m m e r and associates (12) made it possible to ascertain the grams of soaps as well as free f a t t y acids and neutral fat present per 100 g. of fecal d r y matter. The original method gives values for neutral f a t and the total f a t t y acids present as fecal soaps and free f a t t y acids. In the modification an additional titration was p e r f o r m e d on a portion of the dried fecal sample, which was refluxed without the addition of ItC1 to avoid the liberation of the f a t t y acids present as soaps. The value obtained f r o m this additionM titration was taken as the original free f a t t y acids present in the fecal sample. The difference between this vahie and the one obtained after refluxing with acid was taken as the original soaps present in the seraph t of dried feces. I~ESULTS AND DISCUSSION
Fecal characteristics. As mentioned in the earlier report (1), m a r k e d differences in the eonsisten(,y of the feces were f o u n d amoJ~g the various d i e t a r y rogimes. The mean daily fecal excretion values in '['able 1 provide a reasonable TA BLE 1 Fecal o u t p u t by calves on the v'~rious diets
Group l II ]II IV V VI Villi VIII
IX
Diet Corn oil filled nfilk ( p r e p a r e d weekly) Corn oil filled milk (prep:trod daily) l l y d r o g e n a t e d corn oil filled milk L a r d filled milk B u t t e r oil filled milk Fortified skimmilk Fortitied skimmilk plus corn oil Fortified skimmilk plus h y d r o g e n a t e d soybean and cottonseed oil blend Skimmilk
Wet basis.
No. calves
No. tri'ds
Me.m dry matter intake
M e a n daily fecal excretion"
Standard error of m e a n
I0
41
(g.) 829.8
(.q.) 1,170.3
-4- 22.8
4
16
839.1
477.2
-4- 15.1
3 4 3 '2
12 10 12 6
1,161.2 ],256.6 896.8 2,031.8
698.3 557.6 469.2 1,772.8
-4- 23.9 -4-20.6 ~__ 20.6 +--319.7
5
22
1,689.1
],398.1
+
2 2
8 11
2,063.5 667.0
1,428.4 907.8
-4-162.3 +- 55.9
69.0
1564
R . S . ADAMS ET AL
indication of the amount of d i a r r h e a present a m o n g the various groups. The fecal output of subjects in groups with feces of a p p a r e n t l y n o r m a l consistency (Groups I I - V , inclusive) was somewhat greater t h a n t h a t r e p o r t e d b y B l a x t e r and Wood (4) in their s t u d y of diarrhea in calves. Animals fed corn oil filled milk p r e p a r e d weekly were afflicted with a severe diarrhea. I n some trials, as much as 3,000 g. of feces were collected daily f r o m individuals in this group. Tocopherol supplementation of the corn oil filled milk diets did not reduce the severity of the diarrhea. Calves receiving a corn oil filled milk p r e p a r e d daily in an a t t e m p t to reduce its toxicity voided a considerably smaller a m o u n t of feces. H y d r o g e n a t i o n of the corn oil g r e a t l y reduced the amount of feces excreted daily. Similarly to Group I, the feces of animals on the fortified skimmilk plus corn oil diet were extremely loose. Although the mean daily excretion of feces was greatest for the fortified skimmilk group, there was a large amount of variation in the values for t h a t group, as shown b y the s t a n d a r d error of the mean. Considerable v a r i a t i o n in the consistency of the feces was also noted in the trials of animals in Group I X . The m a r k e d fecal excretion b y calves fed the various fortified skimmilk diets undoubtedly was influenced to some extent by the a b n o r m a l l y high solids content of these rations. The factors responsible for the relatively large fecal output b y the skimmilk group are not a p p a r e n t . I t is interesting to note, however, t h a t I o w a workers (13) recently r e p o r t e d that diets high in concentrations of milk solids-not-fat a n d / o r low in f a t are likely to cause diarrhea. Differences in the color of feces were also quite pronounced a m o n g the variot~s experimental rations. The feces of animals on the weekly p r e p a r e d corn oil filled milk diet were characteristically greenish-yellow in color; whereas, those excreted by calves receiving the same diet p r e p a r e d daily were a darker green. The excretions of the lard filled milk group were usually grayish-blue or sometimes clay-like. Subjects on the whole milk, skimmilk, fortified skimmilk, a n d b u t t e r oil filled milk rations voided feces which were the usual brown in color. I n contrast, the fecal excretions of individuals on the h y d r o g e n a t e d vegetable oil filled milks were invariably gray. Differences in bile secretion, intestinal f a t excretion, intestinal flora, and the oxidation-reduction potential of the digestive tract, as well as other factors, m a y influence color of feces.
Digestibility data. The a p p a r e n t d r y m a t t e r digestion coefficients for several of the v,~rious diets may be f o u n d in Table 2. These data were t r e a t e d statistically TABLE 2 A p p a r e n t d r y m;~tter d i g e s t i o n coefficients of t h e v a r i o u s e x p e r i m e n t a l diets
NO.
2~'o.
Diet
c~lves
trials
Corn oil filled milk prepared weekly Corn oil filled milk prepared daily t[ydrogenated corn oil filled milk Lard filled milk Butter oil filled milk
8 4 3 4 3
41
16 12 10 12
Av. dry matter digestion coefficient 81.1 90.4: 89.5 90.9 93.6
EFFECTS
OF FEEDING
FILLED
MILKS
TO CALVES.
II.
1565
according to methods outlined b y Snedecor (11) and D u n c a n (9, 10), to test the differences a m o n g the various t r e a t m e n t means at the 5% level of significance. The average d r y m a t t e r digestion coefficient for the weekly p r e p a r e d corn oil filled milk group was found to be significantly less t h a n t h a t f o r a n y one of the other diets. No other significant differences in a p p a r e n t d r y m a t t e r digestibility were noted among the various diets. The d r y m a t t e r digestion coefficients for calves in the various fortified skimmilk groups were not included in the analyses because of the limited amount of data available for these diets. The significantly greater a p p a r e n t d r y m a t t e r digestibility of the daily prep a r e d corn oil filled milk ration is of considerable interest. Undoubtedly, a large portion of the increase in a p p a r e n t d r y m a t t e r digestibility m a y be associated with the decrease in fecal excretion and degree of diarrhea. As previously noted, the feces of calves in the daily p r e p a r e d corn oil filled milk group were n e a r l y normal in consistency. Blaxter and Wood (4) reported t h a t the a p p a r ent digestibility of fat, protein, and ash drops f r o m as high as 97 to as low as 40% in some instances, as the result of diarrhea in calves. The a p p a r e n t f a t digestibility data are presented in Table 3. The adjusted fat digestion coefficients listed in this table are values adjusted for metabolic fecal fat as follows: The mean a m o u n t of f a t excreted daily by calves in the skimmilk group was expressed as a percentage of the fecal d r y matter. Since the diet of these aninm]s contained only 0.1% fat, it was a ~ u m e d t h a t this value, 5.36%, was indicative of the metabolic fecal fat expressed as a percentage of the fecal d r y matter. Thus, an amount equivalent to 5.36% of the fecal d r y m a t t e r was deducted from the total fat excretion in calculating the adjusted f a t digestion coefficient for calves receiving the sew~,ral filled milks. The data used in determining the metabolic fecal fat excretion were derived f r o m 11 digestion trials conducted with two subjects in the skimmilk group. The finding that the metabolic fecal f a t excretion by calves in this group a m o u n t e d to 5.36% of the fecal d r y m a t t e r is in v e r y close agreement with the 5.30% value obtained under similar conditions by previous workers (5). More recently, C u n n i n g h a m and Loosli (7) re,ported values of 3 . l - 5 . 8 % when the metabolic fecal fat excreted b y calves on a fat-free purified diet was expressed as a percentage of the fecal d r y matter. A?l analysis of variance of the u n a d j u s t e d fat digestibility data indicated that significant diffcre, nces existed among the various diets. F u r t h e r statistical t r e a t m e n t revealed that the digestibility of b u t t e r oil was superior to that of tile fats in each of the other diets studied. I n every ease this superiority of b u t t e r oil was significant at the 1% level. No other significant differences in unadjusted fat digestion coefficients existed among the remaining diets. Signifieant differences were f o u n d to exist also among the adjusted fat digestion coefficients for tile various rations. The digestibility of butter oil was again demonstrated to be significantly greater (1% confidence level) t h a n that of the fats in the other diets. No other significant differences were noted a m o n g the remaining groups. ]it shouht be pointed out that both the highest and lowest f a t digestion coefficients for the over-all experiment occurred in the weekly
TABLE Summary
Diet
of apl)arent
No. calves
fat digestibility
No. trials
3
data for calves on the various experimental
Av. unadjusted fat digestion coefficient
Av. adjusted " fat digestion coefficient
diets
Free fatty aelds
Soaps
Neutral fat
(av. % fecal fat) C o r n oil filled milk prepared weekly C o r n oil filled m i l k prepared daily Hydrogenated c o r n oil filled m i l k L a r d filled m i l k B a t t e r oil filled m i l k Skimmilk
8
32
84.3
88.3 +__ 0.304
22.9 _ 0.53 b
4 2 . 8 ~ 0.85 b
34.3 ___ 0 . 4 8 b
4
14
88.5
90.2 _4- 0.36
1 2 . 7 __+ 0.87
57.7 + 1.20
2 9 . 6 __+ 0.73
3 4 3 2
10 7 9 8
88.2 S7.1 95.1 --
90.3 _+ 0 . 5 4 88.9 ~ 1.06 96.8 _+ 0.26 --
25.7 20.2 25.3 24.2
53.7 52.6 34.3 26.6
20.6 27.2 40.4 49.2
" Values adjusted for metabolic fecal fat as noted in text. ~' M e a n _+ s t a n d a r d e r r o r o f t h e m e a n .
_ 2.26 _ 2.85 ___ 2.62 _ 3.23
~__ 2.22 ~__ 2.99 _.+ 1.70 ~ 2.92
+__ 1.23 -4- 0.79 ___ 1 . 6 9 ___ 2 . 5 7
EFFI~CT8 OF FI~;FDING ~ ] L L E D
M I L K S TO CALVES.
1I.
1567
p r e p a r e d corn oil filled milk group. No consistent effect of collection period upon fat or d r y matter digestibility was observed in any of the groups. I t is interesting to note that the apparent digestibility of the hydrogenated corn oil did not differ significantly from that of corn oil or lard. The melting point range of the hydrogenated corn oil used in this investigation was 4 5 48 ° C. Studies reported by Crockett and Dcuel (6), as well as by others, have indicated that the critical melting point range above which there appears to be a marked decrease in fat digestibility is 50-55 ° C. The a p p a r e n t digestion coefficients for hydrogenated corn oil obtained in this experiment compare favorably with those obtained by Augur, Rollman, and Deuel (2), when rats were fed hydrogenated cottonseed oil with a similar melting point. They are, however, somewhat greater than those for hydrogenated soybean oil as reported for calves by de Man (8). Tile digestibility of lard in this study is somewhat less than that reported by Crockett and Deuel (6) for the rat, but somewhat greater than the 82.8% figure obtained by Cunningham and Loosli (7), when calves were fed a purified diet. The adjusted digestion coefficient of 96.8 for b u t t e r f a t is somewhat greater than the 93.0 figure for calves obtained similarly by earlier workers (5). However, both the adjusted and unadjusted digestion coefficients for b u t t e r f a t are in good agreement with values reported by de Man (8) as well as by Blaxter and Wood (3). It should be pointed out that the daily preparation of the corn oil filled milk did not significantly increase the digestibility of this highly unsaturated vegetable oil, although d r y matter dig~stib.ility was sigmifieantly improved, as noted earlier. The partition of fecal fats is also' presented in Table 3. The finding that a considerable portion of the fecal fat is present as soaps is in agreement with previous results obtained with rats (2, 6) and calves (3). Thus, it appears important that fat determinations be made on acidified samples of fecal d r y matter in fat digestibility studies, in order to include fecal soaps in the total fat excretion. It should be noted that a larger portion of fecal fat was excreted as soaps than as neutral fats and free f a t t y acids combined, by animals on the hydrogenated corn oil diet. Angur, Rollman, and l)euel (2) also showed this to be true in digestibility studies of hydrogenated cottonseed oil with rats. This same pattern of h~;al fat excretion occurred in the daily p r e p a r e d corn oil and lard filb,d milk groups, and was closely approximated b y animals on tim weekly prepared corn oil filled milk ration. Blaxter and Wood (4) obtained similar results with calves fed margarin('. A marked d~,~'rease in the percentage of fecal fat excreted as free f a t t y acids occurred when the corn oil filled milk diet was prepared daily. Undoubtedly, a portion of this decrease is associated with the abatement of the severe diarrhea in this group. Blaxter and Wood (4) demonstrated that a large increase in f a t t y acid excretion results in ~,alves afflicted with diarrhea. Regarding the partition of the metabolic fecal fat as listed under the skimmilk diet in Table 3, the percentage of fecal fat excreted as free f a t t y acids closely agrees with the value obtained by Cannon, Espe, and Waide (5). The percentage of metabolic :fecal fat present as neutral fat, however, is approximately 10% greater than in
1568
~. s. ADAMS ET AL
the results reported by these workers. On the other hand, the portion of metabolic fecal fat present as neutral fat in this investigation is in agreement with the results obtained by Cunningham and Loosli (7). It is interesting to n o t e t h e s i m i l a r i t y b e t w e e n t h e f e c a l f a t p a r t i t i o n o n t h e b u t t e r oil f i l l e d m i l k diet and that of what has been assumed to be metabolic fecal fat, as excreted by subjects in the skimmilk group. ACKNOWLEDGMENTS This research was supported in p a r t by a g r a n t from the National Dairy Council on behalf of the American Dairy Association. The authors wist~ to express their appreciation to D. E. Jones and R. L. Northrup for technical assistance. REFERENCES (1) ADAMS, R. S., GULLICKSON, T. W., GANDER, ft. E., AND SAUTTEI~, J. H. Some Effects of Feeding Various Filled Milks to Dairy Calves. I. Physical Condition and Weight Gains, with Special Reference to Low-Fat Rations. J. Dairy Sc~., 42: 1552. 1959. (2) AUGER, V., ROLLMAN, H. S., AND DEUEL, H. H., JR. The Effect of Crude Lecithin on the Coefficient of Digestibility and :Rate of Absorption of Fat. J. Nutrition, 33: 177. 1947. (3) BLAXTER, 1(. L., ).Nl) WOOD, W. A. The Nutrition of the Young Ayrshire Calf. 5. The Nutritive Value of Cow's Whole Milk. Brit. J. Nutrition, 5: 1. 1952. (4) ]~JLAXTER, ]((. L., AND WO01), W. A. Sonic O1)servations on the Biochemical and Physiological Events Associated with ])iarrima in Calves. Vet. Record, 65: 896. 1953. (5) CANNON, C. Y., ESPE, 2). L., AND WA]DE, J. B., JR. The Relation of Dietary F a t and F a t Derivatives in the Feces of Young Dairy Calves. Proc. Am. Soc. Animal Prod~lction, 1932: 85. ]933. (6) CI~OCKETT, M. E., AND DEUI.~L, ]-[. J., JR. A Comparison of the Coefficient of Digestibility and the ]late of Absorption of Several Natur'Jl and Artificial Fats as Influenced by Melting Point. J. Nutrition, 33: ]87. ]947. (7) CUNNIN(]IIAM, H. M., AND LOOSLI, J. K. The Effect of F a t Free Diets on Young Dairy Calves with Observations on Metabolic Fecal F a t and Digestion Coefficients for Imrd •rod Hydrogenated Coconut Oil. J. Dairy S(:i., 37: 453. 1954. (8) ])EMAN, T~I. J. Kalveropfokproef Met G(q~ydeerde Soya-Olic. Tijdschr. l)i~rgeneesk, 75: 175. 1951. (9) DUNCAN, D. ]~. A Significance Test for Differences Y~etween Ranked Treatments in an Analysis of Variance. Virginia J. Sci., 2: 171. ].(}51. (10) DUNCAN, D. B. On the Properties of the Multiple Comparisons Test. Virginia J. Sci., 3 : 49. 1952. (11) SNEr)~;eO~, G. W. Statistical Methods. 4th ed. The Iowa State College Press, Ames, Iowa. 1946. (12) VAN l)ng KAMMER, d. ~[[., HUININK, H. TEN ]~., AND WEYERS, H. A. R~pid Mc~itod for Determining F a t in Feces. J. Biol. Chew., 177: 347. 1949. (13) WING, J. M., JA(;OmUON, N. L., AN~ AH,EN, l~. S. The Effect of Various Restricted Diets on the Growth and on Certain Blood Components of Young ])airy Calves. J. Dairy Sci., 38: 1006. 1.(}55.