80 RICHARDSON : CHEMICAL ALTERATIONS IN GREEN IX.-Notes o n the Chemical Alterations in Greert Fodder during its Conversion into Ensilage. By CLIFFORD RICHARDSON, Assistant Chemist U.S. Department of Agriculture. IN recent numbers of the Journal of the Chemical Society, Professor Edward Kinch and Dr. 0. Kellner have published some observations on the changes which take place in the nitrogenous constituents of fodders. Analyses which I have made during the past two years throw additional light on the subject., and are, although somewhat incom- plete, presented here. Professor Kinch found that 55 per cent. of the total nitrogen pre- sent in the ensilage of grass was of a non-albuminoid nature, whilst in the original grass only 9 per cent. was of this form. In the case of mangel leaves Kellner found that of the total nitrogen present 27.8 per cent. in the original leaves, 45.5 per cent.in the ensilage, and as much as 59.7 per cent. in the ensilage preserved in stoppered jars was non-albuminoid. In the ensilage of maize I have found :-FODDER DURING ITS CONVERSION INTO EXSILAGE. 81 Per cent. of total nitrogen as non-albumino’id. Original stalks. ..................... 21 -2 Ensilage No. 1.. .................... 44.6 Ensilage No. 2 ...................... 49.6 Dried fodder ...................... 15% Ensilage from young maize .......... 53.3 Ensilage from older maize.. .......... 47.1 These results with grass, mangels, and maize show that in the conditions existing in silos a large portion of the albuminoids are converted into non-albumino’id nitrogenous substances, whilst in the ordinary drying of fodder EO such change seems to take place.Kinch and Kellner both found that a portion of the nitrogen of the original plant was lost or did not appear in the analysis. With grass it was 13 per cent.; with mangels 27.8 per cent. in the unpressed ensilage in the jar, and as much as 59.8 per cent. in that in the open silo. The latter high percentage is doubtless due to the carrying away in the expressed juice of much soluble nitrogenous substance, I am in possession of but few quantitative data in regard to maize, but have found that in the samples of ensilage which I have examined there is a comparatively large amount of ammonia combined with the acids produced by fermentation, acetic and lactic, which o€ course is lost in drying the specimens for analysis.In the results, this pro- duces an apparent loss of nitrogen in the ensilage itself: it is at any rate a loss of nitrogen of nutritive value. A quantitative deter- mination of the ammonia salts in one specimen of ensilage (Serial No. 1693) gave the following results :- Weight of ensilage taken ............ 4000.0 Equivalent to dry substance .......... 620.0 Weight of NHICl 8.660 Per cent. of nitrogen from dry substance 0.366 .................... Equivalent to nitrogen. ............... 2.266 Equivalent to alhumino’ids ............ 2.287 That the nitrogen was in the form of ammonia and not a more complex amine was proved by the following analysis :- Weight of ammonia salt taken ...... Weight of platinochloride found .... 0.1000 0.4105 Equivalent to NHaCl ..............0.0990 This ammonia salt, probably acetate, would be lost in the process of drying, and produce a corresponding deficit in the relative pel’- centage of nitrogen in the analysis. In fact, a loss of 2.29 per cent. of nlbuminoids iu the dry substance of an avemge maize stalk, con- VOL. XLVII. G82 RICHARDSON : CHEMlCAL ALTERATIONS IN GREEN taining 7.50 per cent., amounts to a little moi-e than 30 per cent. of the total nitrogenous substance in the plant, and t o about the loss which Kellner found in his experiments with mange1 leaves not under pressure. The remaining non-albuminoiid nitrogen is probably largely of an amide nature, as, like Kellner, I have found very small amounts of peptones.Attempts, however, to separate any amides in a crystal- line condition have resulted in obtaining nothing but a syrupy nitro- genous substance. The loss of nitrogenous substance from ccnversion into ammonium salts and decomposition on drying is relatively compensated for by the large loss of carbohydrates, so that in the analytical figures neither loss is prominent, and they are at first glance deceptive. The control of course lies in an absolute knowledge of the weight lost by the fodder in the silo, or in the relative increase in one of the constituents which is less liable to change, as for instlance the ash. But as Kellner has shown, pressure is very liable to remove the soluble part of the ash in the juice expressed, and thus entire dependence cannot be placed on this element.An interesting experiment was carried or, at the New Jersey Experi- ment Station in 1881, in which the ash served as a basis for calcula- tion without apparently vitiating the results. Ten tons of green fodder “corn” was divided into two lots on September lst, one half being stacked in the field, and the other packed in a silo of the capacity of 12 tons, after being cut in short lengths. At the same time, a camrefully selected sample of the green fodder was prepared for analysis. In November, 1200 lbs. of the dried fodder was run through a cutter, and an average sample prepared for analysis. On the 23rd of December a specimen of the ensilage was selected. The composition of the three substances was as follows :- Green stalks. Dried stalks. Ensilage.Original substance- Water .......... 75.00 39.37 74.50 Bsh ............ 1-58 4-63 1.97 Fat.. ............ 0.22 0.66 0.27 N.-free extract.. .. 15.60 32.85 13.58 Crude fibre ...... 6.35 18.65 7.92 Crude albumin.. .. 1.25 3.84 1.76 100~00 100~00 100.00FODDER DURING ITS CONVERSION INTO Dry substance- Ash ............ 6.32 7.64 Fat ............ 0.88 1.09 N.-free extract.. .. 62-40 54.18 Crude fibre ...... 2-5-40 30.76 Crude albumin.. .. 5.00 6-33 ENSILAGE. 83 7.71 1.06 53-24 31-07 6-92 ~~ 100~00 100*00 1u0.00 As 100 lbs. of the dry matter of the green fodder contained 6.32 lbs. of ash, the amounts of the other constituents corresponding with this weight of ash and therefore with each original 100 lbs. of stalks, were calculated in order to show the absolute loss of each constituent.Green stalks. Dried stalks. Xnsilage. Ash.. .................... 6.32 6.32 6.32 Fat ...................... 0.88 0.90 0.86 N.-free extract ............ 62.40 44.82 49.64 Crude fibre.. .............. 25.40 25.44 25.49 Crude albumin ............ 5-00 5.24 5.67 Total weight of dry matter to 6.32Ibs. of ash .......... 100*00 82.72 81.98 The loss in t>his case is seen t o fall upon the carbohydrates entirely, and to be as great for the dry fodder as f o r the ensilage. There is an apparent slight increase in albuminolds, which can be explained by the fact that the calculation is made as if no ash had been lost by being dissolved or expressed in the two preserved samples. This is remark- able as showing that in this case, quite at variance with other instances, the nitrogen suffered little o r no loss, and that in fact the whole loss fell upon the carbohydrates.This may he due t o the short time during which the maize was in the silo, as all samples which I have examined have not been taken o u t for several months. These results at least show how varied the conditions are and how unsafe it is to generalise from any one experiment. This point is made evident in the analyses appended t o this paper. As to the nature of the fermentation and the proximate principles involved, I have made some observations. In but one sample out of many examined has any trace of Xaccharomyces been found, As a rule the juice expressed from the fresh ensilage is swarming with Bacillus subtilis, together with some species of Bacterium and Micro- coccu,s.No signs of the ordinary lactic or viscous ferment have been observed. The fermentation does not appear therefore to be of an alcoholic nature, or similar to any with which we are well acquainted. Analyses show the presence of an insufficient amount of alcohol, gum, G 284 RICHARDSON : CHEMICAL ALTERATIONS IN GREEN or free acid for any of the usual forms of fermentation, as can be seen from the following determinations. Alcohol was never absent in any of the ensilages experimented on, but in all was present in such small quantities as to be distinguished only by the iodoform test. Lactic acid has always been detected, but never in large quantity. Acetic acid is the chief acid of the ensilage. The relative proportion of the two acids varies largely, the following being a few deter- minations :- Serial No.1003 ........ 1004 ........ 1500 ........ 1501 ........ 1502 ........ 1677 ........ 1693 ........ Per cent. in ensilage of f - - y - - b - Y Lactic acd. Acetic acid. traces 2.12 0.15 1.59 0.52 0.80 0.24 undet. 0.26 1.02 0.13 undet. 0-11 undet. Total as acetic acid. 1840 .............. 2.40 1841 .............. 1.42 The whole amount of acid present was from 1 t o over 2 per cent., the lactic acid not rising above six-tenths of a per cent. The presence of lactic acid was determined by expressing the juice from a specimen of ensilage, distilling off the volatile acetic acid by repeated distillations, and neutralising the residue with zinc carbonate. The crystals of zinc lactate which were obtained on evaporation were crystallised and analysed.They contained :- Analysis. Theory. Water .......... 18.46 18.1 8 ZnO ............ 26-11 27.27 Gum or similar substance is not formed in any large amounts in the silo. Unchanged sugar has been found in the juice expressed from two ensilages, but as a rule it completely disappears. No. 1003 contained 2.40 per cent., No. 1004 1.07, and No. 1541 0.76 per cent. of reducing sugars. The fibre is of course increased in relative amount, but absolutely it is probable that it is but slightly altered. In Table I (pp. S5-S6) analyses of a number of ensilages are presented together with those of a dried fodder and several green stalks atFODDER DURIKG ITS CONVERSION INTO ESSILAGE. 85 0 m e t-* * - i- E- - .- i i c p p T l .- F 10 9 _ _ _ ~ L- - 1' yyl?FFl- IS -#--mL-cI 0 o*o*--# h X - 4 3 L 0 3 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . r . . . . . - . . . . . * . . . . . . , . . ' X . . . : : , : :.5 : . . : i2 i g . . : s : s I : ; i 5 .a 0) . : i0) :- 3 ; i : E a " g ; i izga i U'aG? 2 % :*?gW ';d ,% d ,-5 I x .gs 4 B 2 c z i; .. . . . . . . . . . . . . . .- c i m o l m t - ??P"W U a w m t - 0 * m * t - - o t o t - CQr-mU2.m mp9y.F -#m . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . * . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . :: : : : : * . . a . 3 : ; : : a ; ; i 13 : :a :- 2 .Q I -TABLE I cont i?aued.-A?zatyses of Maize Silage and of Maize Foddey. ---PA--- Original substance- Water ...................................................................... Ash .......................................................................... Oil, R.C................................................................. Nitrogen-f. ee extract .................................................. Crude fibre ............................................................ N x 6 2 5 = "albuminoids" ., .................................... Dry substance- Ash .......................................................................... Oil, gLc. ................................................................. Nitrogen-free extract ................................................. Crude fibre .............................................................. N x 6.25 = " albuminoids" ...................................... In the cthcr rxtract- In the nit rogen-free extract- Non-volatiitn free acid ...............................................Sugars, &., sol. in 80 prr crnt. alcohol; sol. in water .. Substances, 601. in 80 per cent. aIeoho1; insol. in watcr .. GUUl, R.C. ................................................................ Nitrogen ................................................................. Non-albnxninoid nit,rogen,. .......................................... Pcr cent. of nitrogen as nun-alburninoid ....................... I n the " albuminoids "- I n the original substance- Acetic acid ............................................................ Lactic acid .............................................................. Juice per cent ......................................................... Sugars .................................................................... Solids ...................................................................... Sililge.1 Silage. 16ii. 1 1693. (Blair.) I (Blair.) I _--_____ 81 67 1'04 1-04 1.09 I 0-79 10.17 8 '47 1'39 1 1.34 100'00 ~ 100-00 4'64 1 3'86 __I-___.__ 5 '68 5 .96 55 '50 25 -30 7 '56 100 .oo - -- 0 .i'2 10'59 2 '90 2.91 1 '21 0 '54 44 '6 I 0 *13 - - I Silage. 1652. (Young.) 82 '61 1 .14 1-01 9 2 1 4 '96 1'04 100.00 Silage. 1653. (Old.) --- 78 *62 1-21 1 '46 11.39 5 -80 1 -52 100 '00 6 '68 6 3 9 5 'oa 5 '84 54 -64 53 '08 8 '67 5 9 7 24 '93 28 .:ia p___--__ 100~00 1 100'00 ____---- o -72 l - 5 '68 6 .8? 63 '26 27.12 7-12 100 .oo ---_ 8 '34 - - 3 '66 - - 3'58 I - , - 1-14 0 *54 5 i '1 Dried fodder. 1654. 27 -25 2 .I2 2 3 3 50 -02 1 2 '7.5 5.33 100'00 - --.-- 1-18 0 '18 15 '6 Green fodder.It;"<. St ,lkq for 1677 and 1693. 85 'SF 1 ' X i 0.58 7 .2i 2.48 1 9 0 I00 '00 -- ---- 13.31 3 'il 51 -77 17'67 13 '54 100'00 --- Stalks. Egyptian maize. Y onng anthers just out. -- 81 *90 1 'I1 0 4.1 9 -25 3.04 1.26 100 .oo -_-- 7 *39 2 '89 61 '25 20.15 8 '32 100 *oo -I_---FODDER DURING ITS CONVERSION INTO ENSILAGE. 8'7 various stages of development. From them, much may be learned as to the variations to be expected. The composition of the stalks of green maize is subject to very wide variations, as may be seen by the few analyses quoted. This then is a primary cause of differences in the composition of the ensilage, and, although it is due largely to the period of growth a t which the stalks are cut, yet there are often marked differences in composition in those o f the same stage of development, even in the same field.In a series of analyses of the stalks of Egyptian sugar corn and Lindsay's horse-tooth corn, completed by me, and published in the Beport of the Commissioner of Agriculture for l g 8 l and 1882, these variations are shown, The specimens, the analysis of which are here given, were all from a small plot of carefully selected stalks only a few rods square, and yet, in many instances, they show the largest variations from a regular series. The variations which are found in the composition of the stalks while they are in a condition to be packed in a silo, that is to say, from the appearing of the tops until the grain is well formed, are included within the following limits :- Highest. Lowest.Water in green substance 91.60 79.10 Dry substance.. ........ 20.90 8.40 Ash .................. 9.72 3-54! Oil .................. 3.43 1.68 Crude fibre.. .......... 31.29 21.56 Albuminoids .......... 11.53 1.67 Percentage of nitrogen as non-albumino'id ...... 70.4 18.0 These limits, together with a study of the individual analyses, serve to show the primary cause of the differences in ensilages. As t'o variations produced by other causes, illustrations are found in the samples numbered 1652, 1653, 1677, and 1693. Numbers 1652 and 1653 are analyses of ensilage from young and old stalks. The younger would naturally contain more ash and albuminoYds as it went into the silo, but the nitrogenous substances would be in such a condition, from the greater amount of soluble nitrogen, as to make them more liable to conversion into ammonium salts. This is the case with No.1652. I n it, the ash is higher than in the older speci- men, but the nitrogen is lower from a greater loss. The relative t-lmount of non-albuminold nitrogen is also lower in the younger than in the older sample. I n other respects, they do not differ largely, although one was from stalks on which the ears were well formed, and Carbohydrates .......... 69-40 59.60TABLE 11.-Egyptian Sugar Corn (Maize), plaated A p d 30th, 1881. cx, 'x, ------- D eve lopme 11 t- Height, in feet .................... Diameter, in inches ................ Total weight, in grams.. ............ Weight of stalk.. .................... leaves. .................. .. top ..................... Per cent. of stalk in whole plant.. .... .. ear.. .................... Dry substance- Aih .............................. Criide fat. ........................ Nitrogen-free extract. ............... Crude fibre.. ...................... Crude albumin .................... T o t a l nitrogen .................... Per cent. of nitrogen as non-albumino'id Water. ........................... Ash .............................. Crude f a t . . ..................... Nitrogen-free extract. ............... Crude fibre.. ...................... Crude albumin .................... Non-albumino';d nitrogen. ........... 0 ri gina 1 subs t a n ce- 526. July 5th, top just out. 7.0 1 -2 12442 ' 0 960 .O 216 *O 66 *O 77 *3 - 9 9 2 2 *58 50 -60 27 *01 10 *09 1 *61 0 *99 61 *4 91.60 0 *83 0 *21 4 -25 2 *27 0 -85 540.July l l t h , anthers not out. -- 7 -2 1 '2 1097 * 0 585 -0 468 '0 44.0 53 '3 - 9 *09 1 '80 51 '29 26 '38 11.53 1 '85 1 '27 68 -6 91-30 0 *79 0 .16 4 * d k 5 2.30 1 '01 547. July lltli, anthers out, filling. 8 -5 1-1 1037 *O 5 -42 430 -0 35 -0 52 *1 - 6 -53 1 *68 52 *41 31.29 8.09 1 -29 0.85 65 .9 88 *30 0 9 6 0 -19 6 14 3 -66 0.95 Stalks. 562. J u l s 18th. --- 9 -0 1 -2 1378.0 613 .O 651 *O 26 *O 83 -0 44 -9 5 *60 3.48 60.45 26 -04 4 '43 0 -71 0.31 43 -7 85 *90 0 *79 0 *49 8.53 3 *67 0 -62 576. July 25th, silk out. 9 . 2 1 ' 2 1642 -0 575 0 737 '0 78 *O 252 *O 35 -0 4 -81 2 -7; 61 *53 26 '09 4 *85 0 *78 0 -39 50-0 83 '20 0 -81 0 *46 10 *34 4 -38 0 *81 580. August 1st) our formed.10 -2 1 *6 1722 -0 636 *O 86 2 '0 32 -0 190 0 37 -0 5 -87 2 '35 66 "72 22 -07 2 -99 0 '48 0 *29 60 *5 80 *20 1 *16 0 -47 13 -21 4 '37 0 -59 694. Axgust 8th. 10 *o 1 *2 1068 *O 593.0 300 *O 9 .o 166 .O 55 -5 4 -97 2 el0 67 -37 21 '82 3.74 0 -60 0 -33 55 *o 80 *60 0 -96 0 *41 13.07 4.23 0 -73FODDER DURING ITS CON'VERSION INTO ENSILAGE. 89 the other from stalks on which the ears had made no appearance. In comparison with the ensilages, the dried fodder, No. 1654, from stalks of the same field as the old ensilage, No. 1652, shows several advan- tages. As has been already remarked, its nitrogenous constituents have not suffered so much change, only 15.6 per cent. being in a non- albumindid form as compared with 53.3 and 47.1 per cent. in the ensila'ges. A smaller loss of carbohydrates has left the relative per- centages of fibre, fat, and ash, low; and the evidence points to the fact that the stalks must have dried rapidly and with few changes, furnishing a fodder of much better composition than that analysed in New Jersey.In Nos. 1677 and 1693, we have analyses of two specimens of en- silage taken from the same silo within a few days of each other. One has lost more ash and nitrogen than the other, and this is due probably to greater pressure on the first sample, which had lost expressed juice carrying with it nitrogenous matter and ash. In other respects they are much alike. The variations in composition, which are found among the analyses of ensilage, lie within the following limits :- Water ................ Ash .................. Oil .................. Carbohydrates ........ Fibre ................ Albumino'ids (N x 6.25). . Per cent. of nitrogen as non-albuminoid ...... Dry substance. ......... Ash .................. Oil ................... Carbohydrates ........ Fibre ................ Albumino'ids .......... Or for dry siibstance- Highest. 84.80 2.01 1.80 15-37 7.54 2.77 53.3 15.20 8.87 9-12 61.84 28.58 11.97 Lowest. 70.60 0.9 1 0.79 7.75 2.85 1.04 47.1 29.40 5-68 5-08 48.43 18.76 5.97 By comparison with t.he extremes for green stalks, it is seen that the albuminoids are higher in the dry substance of one of the en- silages, No. 1502, than in the dry substance of any of the stalks. The high figures in the ensilage are probably only relative, due to a great loss of carbohydrates and little change of albuminolds. Exact quavtitative experiments are greatly to be desired, in order that we may have some explanation of the interesting changes which undoubtedly occur. Washington, October, 1884.