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UBC Theses and Dissertations

The evaluation of a new fertilizer derived from fish waste McMullan, Margaret Jean 1947

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THE EVALUATION OF A NEW FERTILIZER  DERIVED PROM FISH WASTE By Margaret Jean McMullan A Thesis Submitted i n P a r t i a l F u l f i l m e n t of the Requirements f o r the Degree of MASTER OF SCIENCE IN AGRICULTURE i n the DEPARTMENT OF HORTICULTURE Approved* (Major) ' (Minor) The U n i v e r s i t y of B r i t i s h Columbia A p r i l , 1947. ACKNOWLEDGEMENT The w r i t e r wishes to express a p p r e c i a t i o n to S r . A . F . Baras, Professor and Head of the Department of H o r t i c u l t u r e at The U n i v e r s i t y of B r i t i s h Columbia, f o r his encouragement i n the completion of t h i s experiment; and to extend g r a t e f u l thanks to D r . G.H. H a r r i s , Professor of Plant N u t r i t i o n i n the Department of H o r t i c u l t u r e , f o r h i s valuable suggestions and c r i t i c i s m i n c a r r y i n g out the p r o j e c t . TABLE OP CONTENTS 1. INTRODUCTION (a) The F e r t i l i z e r M a t e r i a l Under Test 1 (b) The Object of the Experiment. . 1 (c) Review of L i t e r a t u r e . 2 2. MATERIALS AND METHODSt (a) General R e m a r k s . . . . . . . . . . . . 3 (b) S o i l s 5 (c) F e r t i l i z e r 6 (d) Equipment. 6 (e) Pot Setup f o r P e r c o l a t i o n 7 (f) Examination and A n a l y s i s of P e r c o l a t e . . . 8 (g) Pot Setup for Plant Growth 10 (h) Weighing of Plant M a t e r i a l . . . . 10 3. OBSERVATIONS AND RESULTS« 10 4. DISCUSSION OP RESULTS 22 5. CONCLUSIONS,.^ 29 6. SUMMARY, 31 7. REFERENCE S« 32 THE EVALUATION OP- A-NEW FERTILIZER  DERIVED FROM PISH WASTE . By Margaret Jean McMallan Abstract Water i n three separate a p p l i c a t i o n s was allowed to p e r c o l a t e through columns of pots c o n t a i n i n g d i f f e r e n t types of s o i l s to which f i s h p r o t e i n a t e had been added only i n the top pot . The volume of water was s u f f i c i e n t to saturate a l l the s o i l and to give a small excess for a n a l y s i s . ' Plants were subsequently grown i n a l l pots and the inoreased growth i n d i f f e r e n t pot l e v e l s was used as a measure of nitrogen r e t e n t i o n . On the whole the f i s h proteinate was well r e t a i n e d , p a r t i c u l a r l y i n heavy s o i l s , but the r e t a i n e d f e r t i l i z e r was more e f f i c i e n t l y u t i l i z e d i n l i g h t s o i l . The tendency of the f i s h proteinate i n i t s present form to absorb moisture from the a i r and to cause the formation of a hard c r u s t , e s p e c i a l l y i n the l i g h t s o i l - , are d e f i n i t e drawbacks, and i t i s concluded that these undesirable features must be overcome before the product should be put on the market. THE EVALUATION OE A NEW FERTILIZER DERIVED FROM FISH WASTE 1. INTRODUOTIOffi (a) The P e r t l l l a e r M a t e r i a l Under Testt In s p i t e of the urgent demand f o r nitrogenous material f o r n u t r i t i o n a l , f e r t i l l z e r , and I n d u s t r i a l uses* a v a i l a b l e sources of n i t r o g e n i n the form of f i s h waste are discarded i n tremendous q u a n t i t i e s every year. One manu-f a c t u r e r i n B r i t i s h Columbia who obtains o i l from f i s h waste by chemical e x t r a c t i o n i s endeavouring to u t i l i z e the nitrogenous residue from t h i s process by c o n v e r t i n g i t to a form s u i t a b l e f o r f e r t i l i z e r . The m a t e r i a l i s d r i e d to a powder and then mixed with dlatomaoeouB e a r t h i n an e f f o r t to overcome the tendency to absorb moisture. The f i n a l product has a r e a c t i o n ranging from pH 6 to pE 10 and a n i t r o g e n content of approximately 8%, The nature of the nitrogenous m a t e r i a l i n the f e r t i l i z e r has not been a s c e r t a i n e d ; i t i s r e f e r r e d to by the manufacturer as H f l s h p r o t e l n a t e H . (b) The Object o f the Experiment! The experiment was c a r r i e d out i n the greenhouse to supplement a test already made (6) and a f i e l d experiment c u r r e n t l y i n progress (5) to determine the f e a s i b i l i t y of using the f i s h protelnate as a commercial f e r t i l i z e r . -The main purpose of t h i s experiment was to obtain information regarding the r e t e n t i o n of the f e r t i l i z e r i n successive s o i l l a y e r s and the a v a i l a b i l i t y of the n i t r o g e n i n d i f f e r e n t s o i l types; and a l s o to observe the degree of l e a c h i n g of the m a t e r i a l , i t s ef fect on the p h y s i c a l nature of the s o i l s , and some of the p r o p e r t i e s of the f e r t i l i z e r i t s e l f * (e) Review of L i t e r a t u r e s Sue to the fact that so l i t t l e was known about the chemical nature of the material under t e s t , the l i t e r a t u r e reviewed was n e c e s s a r i l y r e s t r i c t e d , and dealt only with a method p r e v i o u s l y used to evaluate the r e t e n t i o n of n i t r o g e n i n s o i l s * The procedure used i n this experiment i s an a d a p t a t i o n of the p e r c o l a t i o n method used by Conrad ( 2 ) to determine the degree of r e t e n t i o n by s o i l of nitrogen from a wide v a r i e t y of sources. The method gives semi-quantitat ive r e s u l t s , and provides a very s a t i s f a c t o r y means of determining the retention i n the s o i l of ni trogen from complex organic compounds whose r e a c t i o n with the s o i l oannot be p r e d i c t e d with any degree of accuracy* Conrad's method c o n s i s t e d of p e r c o l a t i n g a s o l u t i o n of the m a t e r i a l under t e s t through a column of three pots and subsequently measuring the growth of p l a n t s i n the top, middle, and bottom pots* By t h i s means he showed that the nitrogen contained i n anionic u n i t s i n compounds was not r e t a i n e d i n the s o i l s tested; that nitrogen contained i n c a t i o n i c u n i t s of the compounds was completely or almost completely r e t a i n e d i n the top pots of t h e i r respect ive columns. Nitrogen i n amphoteric or approximately n e u t r a l u n i t s were unpredictable and t h e i r behaviour v a r i e d c o n s i d e r a b l y . Since the exact chemical nature of the n i t r o g e n f r a c t i o n of the f i s h proteinate was not known, i t was Impossible to p r e d i c t the r e a c t i o n of the material with the s o i l and the consequent degree of r e t e n t i o n . 2. MATiJiRXALS M P IPTHQpst (a) General Remarksi Conrad* s method consisted of p e r c o l a t i n g a s o l u -t i o n of the nitrogenous material through the column of p o t s , but i n the present experiment, due t o the nature and composition of the f e r t i l i z e r mixture* i t was deemed a d v i s -able to Incorporate the f i s h p r o t e i n a t e with the s o i l i n t h e top pot In each column and to percolate only water through the s e r i e s . This method served two purposes* It produced c o n d i t i o n s somewhat comparable to those i n a f i e l d on which f e r t i l i z e r had been broadcast and harrowed i n , and I t prevented the accumulation on the surfaoe of the s o i l of the diatomaceous e a r t h contained i n the f e r t i l i z e r . A f u r t h e r m o d i f i c a t i o n of Conrad* s method was made by c a r r y i n g out the leaching procedure three times i n s t e a d of once. The s o i l was thus subjected to very severe l e a c h i n g , and i t was considered that i f the f e r t i l i z e r was r e t a i n e d i n s p i t e of such treatment, the- degree of r e t e n t i o n most be very good. Furthermore, the comparatively long p e r i o d between the f i r s t leaching and the i n i t i a t i o n of the growing t e s t allowed time f o r the breaking down of the f i s h proteinate i n t o a form a v a i l a b l e f o r plant growth. It was hoped that p l a n t response would be an i n d i c a t i o n of the rate of a v a i l a b i l i t y of the nitrogen as w e l l as an i n d i c a t i o n of the degree of r e t e n t i o n i n the d i f f e r e n t s o i l s . Owing to the large number of percolate samples and to the fact that the object of a n a l y s i n g the p e r c o l a t e was to obtain an i n d i c a t i o n of ranges r a t h e r than exact v a l u e s , i t was decided to make use of a c o l o r i m e t r i c method f o r est imating the t o t a l nitrogen and the n i t r a t e nitrogen i n the p e r c o l a t e * Emmert's (3) method of nitrogen determination was t r i e d and numerous tests were made on the f i r s t p e r c o l a t e , but c o n -s i s t e n t r e s u l t s c o u l d not be obtained and the method had t o be d i s c a r d e d . Sue to the f a i l u r e of t h i s method, b a c t e r i a l growth was pronounced i n a l l percolate samples from test columns, with the r e s u l t t h a t no n i t r a t e nitrogen estimations could be made on these samples. The amount of b a c t e r i a l growth was recorded as being a p o s s i b l e i n d i c a t i o n of the amount of nitrogen i n the p e r c o l a t e * T o t a l n i t r o g e n determinations were c a r r i e d out by another method on a l l the p e r c o l a t e samples from the f i r s t l e a c h i n g , though i t was r e a l i z e d that there might be some e r r o r due to decomposition of the p r o t e i n a t e . The method of nitrogen a n a l y s i s f i n a l l y adopted was a combination of Emmert's (3) d i g e s t i o n prooedure and one of Bray's (1) methods f o r the measurement of n i t r a t e n i t r o g e n . (b) s o n s i In order that the p r o p e r t i e s and r e a c t i o n s of the f i s h p r o t e i n a t e could be studied on a f a i r l y wide b a s i s , s o i l s * of various types found i n the a g r i c u l t u r a l areas of the Lower Mainland of B r i t i s h Columbia were used i n the experiment. These s o i l s lnoluded a heavy c l a y , a f ine loam h i g h i n organic matter, muck, and a l i g h t sandy s o i l . No attempt was made to a l t e r the r e a c t i o n of the s o i l s , nor to add supplementary elements, as i t was considered that such changes would create more problems than they s o l v e d . The four s o i l s used i n the experiment were designated as followst SOIL At u n t l l l e d c l a y s o i l from Sea I s l a n d , c l a s s i -f i e d on the S o i l Hap of the Lower Fraser V a l l e y (4) as Ladner C l a y . SOIL Bt u n t i l l e d s o i l from Langley Towns!te, c l a s s i -f i e d as Custer Loam. SOIL Ct u n t i l l e d Blaok Muck from L u l u Island near Brlghouse. SOIL St Upland Sandy Loam, p r e v i o u s l y c u l t i v a t e d , from the H o r t i c u l t u r e area of the Campus of The U n i v e r s i t y of B r i t i s h Columbia. In taking the samples of u n t i l l e d s o i l s , the cover of weeds and the top two inches of s o i l were removed and the s o i l taken t o a depth of about eight i n c h e s . The s o i l s were r e c e i v e d In f i e l d - m o i s t c o n d i t i o n and a f t e r they had been s i f t e d through a h a l f - I n c h mesh to remove lumps, stones, r o o t s , e t c . , they were then spread on a bench In the greenhouse to dry. They were tnrned and s t i r r e d frequently f o r three weeks, and were then considered to be i n an a i r - d r y c o n d i t i o n . (e) F e r t i l i z e r , The f e r t i l i z e r m a t e r i a l used i n t h i s experiment has already been described on page 1* (d) Bflttlttmeat* Potst 60 f o u r - i n c h pots complete with one-hole corks and "needle-valves* made of g l a s s tubing* served as r e s e r v o i r s . ISO f o u r - i n c h pots were "used to contain the s o i l . A l l pots were painted on the outside with asphalt p a i n t . Canst 180 cannery t i n s * No. 2-§-, painted on the Inside with asphalt p a i n t , were used to catch the excess drainage from pots c o n t a i n i n g p l a n t s * J a r s i 96 four-ounce j a r s were used to catch the percolate and to hold samples f o r a n a l y s i s . Racksi 3 wooden racks with removable side bars were designed to hold double rows of p o t s , as shown i n Plate 1. Chemical 8 1 i n a d d i t i o n to the chemicals s p e c i -f i e d In the a n a l y t i c a l procedures, the fol lowing reagents were made up, Reftgen^ 1» 100 gm. BaS0 4 1 gm. f i n e l y powdered z i n c 10 gm. M n S 0 4 . H 2 0 The reagents should be free from n i t r a t e and n i t r i t e and be ground together forming a nearly white powder. Reagent g: 10/B a c e t i c a c i d made n i t r a t e - f r e e by the a d d i t i o n of a l i t t l e powdered zinc. Reagent St To 50 m l . of Reagent 2 add a p p r o x i -mately 0.25 gm. of a mixture of equal parts by weight of a l p h a - n a p h t h y l -amine and s u l p h a n i l l o a c i d . Plant s i "King of Denmark1* spinach p l a n t s were grown to p r i c k i n g - o u t s i z e i n a f l a t and then transplanted to the p o t s . (e) Pot Setnp f o r P e r c o l a ^ o n , . Columns of pots were set up with three pots per column. Bach pot oontained 300 gm. of a i r - d r y s o i l , except i n the case of SOIL C (Blaok Muck) when only 200 gm. could be put i n each p o t . With' the s o i l i n the top pot of each column was i n c o r -porated 11.2 gm. of f i s h p r o t e i n a t e (7.5 gm. i n SOIL C) so that the nitrogen l e v e l of the s o i l of the top pot was r a i s e d by approximately 0.3J6 on an a i r - d r y b a s i s . The s o i l and the f e r t i l i z e r were thoroughly mixed by shaking the two together i n a c losed can. Test columns were r e p l i c a t e d twelve times f o r each s o i l . S i m i l a r columns of each s o i l without added f e r t i l i z e r were set up to serve as ohecks. Check columns were r e p l i c a t e d three times f o r each s o i l . Before each pot was f i l l e d with s o i l * a pieoe of p o t t e r y was plaoed over the drainage h o l e , and a small handful of washed gravel was dropped i n to hold the p o t t e r y i n place and to provide good drainage and a e r a t i o n . From a r e s e r v o i r pot above each column, tap water was p e r c o l a t e d through the s o i l u n t i l an excess of more than 20 ml. had been c o l l e c t e d i n a j a r set below the column. The water was dripped at a s u f f i c i e n t l y slow rate to prevent the formation of surface p o o l s , but fast enough so that a l l p e r c o l a t i n g was completed w i t h i n 48 hours. No record was kept of the amount of water r e q u i r e d to saturate each type of s o l i . 20 ml. of percolate was saved from each column f o r a n a l y s i s , and the remainder was returned to the top pot . In the oase of t e s t columns, the percolate from each group of three columns was made i n t o a composite sample i n order to reduce the number of nitrogen estimations to be made. The sample of percolate from each oneok column was placed i n a separate storage j a r . The p e r c o l a t i o n process was c a r r i e d out three times, with an i n t e r v a l of 28 days between the f i r s t and second, and 8 days between the second and t h i r d l e a c h i n g s . (f) Examination and A n a l y s i s of P e r c o l a t e i Observations were made on the colour of the p e r c o -l a t e at the time i t was c o l l e c t e d , with the thought i n mind that there might be an a s s o c i a t i o n between the colour of the percolate and the amount of n i t r o g e n leached out of the s o i l * It was remarked above that due to the delay i n f i n d i n g a s a t i s f a c t o r y c o l o r i m e t r i c test f o r t o t a l n i t r o g e n i n the f i r s t leachate, b a c t e r i a l growth was evident i n a l l percolate samples from columns r e c e i v i n g f e r t i l i z e r . An e s t i m a t i o n of the amount of growth was made by v i s u a l examination. The r e a c t i o n of the l a s t p e r c o l a t e was recorded to show the cumulative effeot of l e a c h i n g on the a c i d i t y of t e s t and o;heck s o i l s and to e s t a b l i s h the pH of the s o i l s at the time the growing t e s t was begun. The approximate a c i d i t y was determined by the use of Hydrion paper. A small piece of the paper was placed i n the depression of a spot plate and j u s t moistened with p e r c o l a t e . The colour developed was then compared with the standards p r o v i d e d . D i g e s t i o n p r i o r to the determination of t o t a l n i t r o g e n was c a r r i e d out by Emmert's (3) method as f o l l o w s : 5 m l . of the percolate were placed i n a test tube and evaporated to dryness i n a water bath. To the dried m a t e r i a l was added 1 m l . of A0% sodium chlorate s o l u t i o n without wetting the sides of the tube. The tube was then s l a n t e d and 1 m l . of fuming s u l p h u r i c a c i d (16% S O g ) was allowed to run slowly down the side of the tube with constant shaking of the tube to prevent s p a t t e r i n g . When the r e a c t i o n had ceased 3 m l . of d i s t i l l e d water were added. 1 m l . of the digest s o l u t i o n was then placed i n a clean test tube with 2 drops of phenolphthalein and the s o l u t i o n was n e u t r a l i z e d with 40% sodium hydroxide. S u f f i c i e n t 10% a c e t i c a c i d was added to dispel the red c o l o u r , and then 10 drops more. The volume was made up to 5 m l . with d i s t i l l e d water* Using the n e u t r a l i z e d digest s o l u t i o n f o r t o t a l n i t r o g e n and the percolate f o r n i t r a t e n i t r o g e n , a c o l o r i m e t r i c e s t i -mation of the nitrogen content was made by B r a y ' s (1) method as f o l l o w s ! - 10 -To duplicate 1 ml* samples i n a t e s t tube were added 7 mL of Reagent 2, and then £ ml. of Reagent 1. The tube was shaken immediately f o r exact ly 20 seconds and then 1 m l . of Reagent 3 was added and the tube shaken onoe more. The tubes were allowed to stand f o r several hours or overnight before comparison was made with a s e r i e s of standards prepared by the same method. (g) got Setup for Plant Srowthi The columns of pots were taken down and each pot was placed i n a t i n can as shown i n PLATES I I - I V . One spinach plant was set i n each p o t . The pots were randomised on the greenhouse bench and watered as necessary. Excess water ',• c o l l e c t e d In the t i n cans was returned to the p o t s . A f t e r about eight weeks of growth, when the lower leaves were begin-ning to show c h l o r o s i s , the plants were harvested. (h) Weighing of Plant M a t e r i a l i The tops of plants were weighed immediately a f t e r h a r v e s t i n g . 3. OBSERVATIONS AND RESULTSt (a) ffroperUes of ths F e r t i l i z e r Matftna.it In s p i t e of the a d d i t i o n of diatomaoeous e a r t h to the d r i e d f i s h p r o t e i n a t e , i t s t i l l e x h i b i t e d pronounced d e l i -quescence, and was d i f f i c u l t to handle a f t e r short exposure to the a i r . Thus even weighing the. f e r t i l i s e r i n a beaker presented d i f f i c u l t i e s , as the material absorbed moisture f a s t enough to s t i c k to the sides of the g l a s s . The weighed m a t e r i a l could not be used immediately and was s t o r e d i n g l a s s j a r s with t ight screw caps; at the end of a two months the o r i g i n a l powder had become a hard, unmanage-able mass, and new p o r t i o n s of f e r t i l i s e r had to be weighed out. (b) E f f e c t of the F e r t i l i s e r on jfee S o ^ i At the time of the second l e a c h i n g i t was observed that a l l s o i l s In columns which had received f e r t i l i z e r were considerably caked on the s u r f a c e . In the case of SOIL D (Upland Sandy Loam) the s o i l was quite impervious to water u n t i l the c r u s t had been broken. This severe caking e f f e c t i n SOIL D was s t i l l evident at the time of t r a n s p l a n t i n g s e e d l i n g s ; i n the other s o i l s the e f f e c t had been e l i m i n a t e d almost e n t i r e l y by successive, l e a e h i n g s . (c) P e r c o l a t e , Oolonrt The r e s u l t s of these observations are tabulated i n TABLE I . B a c t e r i a l Growtht TABLE II records an est imation of the amount of growth i n percolates from the d i f f e r e n t s o i l s f o l l o w i n g the f i r s t l e a c h i n g . pHt The r e s u l t s of the a c i d i t y estimations on the t h i r d p e r c o l a t e are shown i n TABLE I I I . Nitrogen Content* In TABLE IV Is shown a summary of the estimations of t o t a l n i t r o g e n and n i t r a t e nitrogen In the percolate c o l l e c t e d from a l l columns of p o t s , and the increase i n the t o t a l nitrogen leached from the s o i l s as a r e s u l t of the a d d i t i o n of f e r t i l i z e r * Qrowtht The comparative s i z e s of p l a n t s i n the check, top, middle, and bottom pots are shown i n the photographs i n PLATES I I - I V . P l a n t s i n SOIL C (Black Hack) made very l i t t l e growth and a f t e r s i x weeks i n the pots showed c h l o r o s i s and s t u n t i n g to such a degree that the experiment w i t h t h i s s e r i e s of p l a n t s was discontinued and no f u r t h e r observations were made* F r e s h Weight* Due to the small s i z e of the p l a n t s , they were not weighed i n d i v i d u a l l y . The twelve plants ' growing In the top pot of each s o i l type were weighed and the average f r e s h weight per p l a n t was c a l c u l a t e d . S i m i l a r l y , the p l a n t s of the middle and bottom pots ware weighed and the average weights found. The three p l a n t s i n the top* middle, and bottom pots of each cheek s o i l were also weighed and the average weight recorded. The average fresh weights per p l a n t are shown In TABLE V along with the Increase i n weight and the percentage increase as a r e s u l t of the f e r t i l i z e r 1 a p p l i c a t i o n * - 13 -sun i r POT SETUP IN RACKS FOR PERCOLATION TEST PLANT GROWTH IN SOIL A (LADNER CLAY) (Check, Top, Middle, and Bottom Pots) - 15 -EMJJ III PLANT GROWTH IN SOIL D (UPLAND SANDY LOAM) (Cheek, Top, M i d d l e , and Bottom Pots) 17 -Colour o f the F i r s t P e r c o l a t e Test Check SOIL A Orange-ye11ow F a i n t yellow SOIL B Almost c o l o u r l e s s C o l o u r l e s s SOIL C L i g h t brown Orange-yellow SOIL D L i g h t brown Very f a i n t yellow - 18 TABLE U B a c t e r i a l Growth i n the F i r s t Percolate Test . Check SOIL A SOIL B SOIL C SOIL 9 KKYt None + S l i g h t ++ Moderate +++ Heavy ++++ Yery Heavy - 19 TAPfcJ Ml pH of the T h i r d Percolate Teat Check SOIL A pH 7.0 pH 6.5 SOIL B pH 5.0 pH 6.5 SOIL C pH 4.0 pH 4.5 SOIL D pH 7.5 pH 7.0 20 -Nitrogen A n a l y s i s of the P e r c o l a t e s Total Nitrogen me. tier l i t r e N i t r a t e Nitrogen me. per l i t r e F i r s t ; Second T h i r d T o t a l F i r s t Second T h i r d SQIL, A Test 76 * 100 15 190 20 10 Check 60 75 10 145 15 15 7 Increase 15 25 .5 11 SPXJ, ,B Test 200 100 50 350 30 12 Check 100 25 10 135 25 15 7 Increase ' 100 75 40 m± SQIL, & Test 200 175 50 425 20 12 Cheek 100 100 25 225 15 15 12 Increase 100 75 25 200 SQJL. P Test 750 250 25 1025 25 15 Cheek 200 25 15 i 240 50 20 12 Increase 550 225 10 785 - 21 -Average F r e s h Weight - Grans Per Plant Top Pot Middle Pot Bottom Pot Test 2.52 2.16 2.56 Check 0.70 ' 1.67 1.87 Increase 1.82 0.49 0.69 Percent Increase 262 ZSL 57-SOIL. B Test 0.86 0.67 0.60 Check 0.30 0.53 0.40 Increase 0.56 0.14 0.20 Percent Increase 187 £& 50 SOIL 0 NQ M T A SOIL D Test 1.90 Check 0.35 Increase 1.55 Percent Increase 444 1.30 1.30 0.35 0.37 0.95 0.93 272 251 Notet TB3T. Average of 12 r e p l i c a t i o n s CHECK-t Average of 3 r e p l i c a t i o n s - 22 -4. DISCUSSION OP RBSULTSi (a) Colour of the F l r s t P e r c o l a t e ! Reference to TABLE I shows that the greatest colour change In the f i r s t p e r c o l a t e between f e r t i l i s e d and u n f e r t i -l i z e d s o i l s occurred i n SOIL D (Upland Sandy Loam) and comparison with TABLE IV shows that by f a r the greatest l o s s of t o t a l ni trogen a l s o occurred from t h i s s o l i * Here the r e l a t i o n between colour change and degree of leaohing seems to end. f o r moderately heavy l o s s e s of t o t a l nitrogen occurred from SOIL B (Custer Loam) and SOIL C (Black Muck) whereas the colour changes were s l i g h t ; and i n the case of SOIL A (Ladner Clay) the colour change was quite pronounced but the loss of nitrogen was comparatively s m a l l . 1 Thus i t seems that the colour of the p e r c o l a t e as a r e s u l t of adding f i s h proteinate to a s o i l i s not n e c e s s a r i l y a measure of the amount of ni trogen c a r r i e d out i n the leachate, p a r t i c u l a r l y i n f i n e - t e x t u r e d s o i l s * Since the nitrogen f r a c t i o n of the f e r t i l i z e r i s not h e l d i n a l l s o i l s to the same degree as are the t o t a l organio c o n -s t i t u e n t s of the f e r t i l i z e r * no d i r e c t r e l a t i o n can be said to e x i s t between l o s s of nitrogen and the colour change induced i n the p e r c o l a t e * (b) B a c t e r i a l Growth i n the F i r s t P e r o o l a t e i The amount of b a c t e r i a l growth i n the f i r s t percolate from the s o i l s r e c e i v i n g f i s h p r o t e i n a t e agrees to some degree with the increased nitrogen i n the p e r c o l a t e * to the extent that In the leaehate from SOIL A (Ladner Clay) the nitrogen oontent was low and growth was slight« and i n the leaehate from SOIL D (Upland Sandy Loam) the n i t r o g e n content was very high and growth was very heavy. Although the amount of proteinate nitrogen was the same In the p e r c o l a t e s from SOIL' B (Custer Loam) and SOIL 0 (Bla'ck Muck), b a c t e r i a l growth was considerably heavier i n the l a t t e r . Since the percolate from s o i l s r e c e i v i n g no f e r t i l i z e r were f a i r l y high i n t o t a l nitrogen i n some cases but showed no growth of b a c t e r i a , i t appears that b a c t e r i a l a c t i v i t y depends on the form In which the nitrogen o c c u r s . However, since the growth i n the leaehate i s not p r o p o r t i o n a l i n a l l cases to the amount of nitrogen derived from the p r o t e i n a t e , another f a o t o r must be present i n varying amounts, and i t i s suggested that organic matter c a r r i e d down i n the drainage water may be the f a c t o r i n v o l v e d . ( c) p H of the T h i r d P e r c o l a t e i No consistency i s apparent i n the ef fect of the proteinate on the r e a c t i o n of the s o i l p e r c o l a t e , B B i t w i l l be observed i n TABLE III that the pH was r a i s e d i n two cases and lowered i n two cases compared with the p e r c o l a t e s from s o i l s which had r e c e i v e d no f e r t i l i z e r . However, since the Hydrion paper method of measuring a c i d i t y i s not s e n s i t i v e to f ine g r a d a t i o n s , i t i s suggested that dif ferences of 0.5 one way or the other should be ignored, and f o r p r a c t i c a l purposes i t can be s a i d that the f i s h p r o t e i n a t e caused no change of - 24 -pH i n SOILS A , 0. and D. The pronounced Increase of a c i d i t y i n SOIL B (Custer Loam) i s apparently associated with b a c t e r i a l a c t i v i t y and the organic matter f r a c t i o n of the s o i l . It w i l l be observed that i n the Black Muck a reduction of pH a l s o occurred* whereas i n the mineral s o i l s the pH showed a tendency to r i s e as a r e s u l t of the f e r t i l i z e r a p p l i c a t i o n . (d) Njtroeen Analyses of the Percolate* Leaching of t o t a l nitrogen from the d i f f e r e n t s o i l s v a r i e d widely. Thus i t i s seen i n TABLE IV that the t o t a l nitrogen l o s t from SOIL D (Upland Sandy Loam) was very l a r g e , whereas the amount l o s t from SOIL A (Ladner Clay) was comparatively s l i g h t , and the losses from SOIL B (Ouster Loam) and SOIL C (Black Muck) were intermediate. The inorease i n t o t a l nitrogen leached i n the three p e r c o l a t e s as a r e s u l t of the a d d i t i o n of f i s h p r o t e i n a t e i s shown f o r purposes of comparison between the d i f f e r e n t s o i l s . These increases i n n i t r o g e n l o s t through l e a c h i n g can be expressed i n simple r a t i o s or "Leaching Factors ' 1 as f o l l o w s : SOIL A (Ladner Clay) 2 SOIL B (Custer Loam) 10 SOIL C (Black Muck) 9 SOIL D (Upland Sandy Loam) 35 It i s i n t e r e s t i n g to note that i n SOIL A (Ladner Clay) the losses of nitrogen from both t e s t and check pots were - 25 -g r e a t e r i n the second than i n the f i r s t p e r c o l a t e . .Apparently the long i n t e r v a l between the leachings caused more of the nitrogen to become soluble and free i n the s o i l . There seems to be no p a r t i c u l a r s i g n i f i c a n c e to the losses of n i t r a t e n i t r o g e n * e s p e c i a l l y i n view of the fact that e s t i -mations for the f i r s t percolate are m i s s i n g . (e) Retention of Nltrogent Conrad i n t e r p r e t s h i s r e s u l t s of the growing t e s t by observing the r e l a t i v e growth i n the top pot compared to growth i n the middle and bottom pots of the t e s t columns, and unless the d i f f e r e n c e s are very g r e a t , he seems to conclude that r e t e n t i o n was s l i g h t . In the present experiment, howevez; three leaohings were c a r r i e d out Instead of one, with the r e s u l t that a l a r g e r p r o p o r t i o n of the n u t r i e n t m a t e r i a l was c a r r i e d down from the top pot to the middle and bottom pots i n both t e s t and check columns. Hence, i t appears that a better est imation of r e t e n t i o n can be a r r i v e d at by f i n d i n g the inorease i n p l a n t growth at each pot l e v e l r e s u l t i n g from the a p p l i c a t i o n of the f i s h p r o t e i n a t e . The increase i s expressed as a percentage of the fresh weight of p l a n t s i n the check p o t s , as shown i n TABLE V. When a comparison i s made on the basis of percentage inorease i n growth, i t becomes evident that response i n the top pots i s considerably greater than i n the middle and bottom p o t s . These data are i n t e r p r e t e d g r a p h i c a l l y i n FIGURE I . To f a c i l i t a t e comparisons between dif ferent s o i l types, a 26 - 27 -"Retention Factor 1 * i s derived by d i v i d i n g the increase i n the top pot by the increase i n the bottom p o t . (The bottom pot* rather than the one showing the lowest percentage i n c r e a s e * was chosen a r b i t r a r i l y f o r the sake of u n i f o r m i t y ) . In i n t e g r a l numbers the r e t e n t i o n of the p r o t e i n a t e i n the d i f -ferent s o i l s i s then shown as f o l l o w s : SOIL A (Ladner Clay) 7 SOIL B (Custer Loam) 4 SOIL 0 (Black Muck) . SOIL D (Upland Sandy Loam) 2 Further examination of the r e s u l t s of t h i s experiment give some i n d i c a t i o n of the nature of the r e t e n t i o n i n the s o i l as well as i t s degree. In the case of the l i g h t s o i l (SOIL D, Upland Sandy Loam) i t appears that the f i s h p r o t e i n a t e was washed down into the middle and bottom pots and i n t o the drainage water c o l l e c t e d during the f i r s t leaohing procedure. This c o n c l u s i o n i s borne out by the pronounced change i n colour of the percolate and the heavy b a c t e r i a l growth. The p r o t e i n a t e held i n the s o i l layers was subsequently broken down at a comparatively r a p i d rate and became a v a i l a b l e f o r plant growth; hence the growth at a l l l e v e l s was very high compared to the ohecks. In. the heavier s o i l s , however, the p r o t e i n a t e was almost completely r e t a i n e d i n the top pot and apparently only a small soluble f r a c t i o n p e r c o l a t e d through the middle and bottom pots and i n t o the excess leachate c o l l e c t e d f o r a n a l y s i s . I f such were the case, the soluble material should - 28 -be more concentrated i n the bottom than in the middle pot* and therefore increased growth should be g r e a t e r in the bottom pot. Such was the case i n both SOIL A (Ladner Clay) and SOIL B (Custer Loam)* as i s c l e a r l y shown i n FIGURE I . Since the proteinate was held very l a r g e l y i n the top pot* growth at that l e v e l was much greater than i n the middle and bottom pots. Due* however* to the slow rate of a v a i l a b i l i t y i n these heavy s o i l s , the increased growth compared to check p l a n t s was much l e s s than i n the l i g h t B o i l . In view of the above observations, i t seems that the f i s h proteinate i s retained i n the s o i l i n i t s o r i g i n a l form rather than as a product of enzyme or b a c t e r i a l decomposition; i n other words, r e t e n t i o n of t h i s material i s a p h y s i c a l r a t h e r than a chemical phenomenon. (f) A v a i l a b i l i t y of the r-e-rtJ* 3,1 sari Although the main object of the experiment was to estimate the degree of r e t e n t i o n of the f i s h p r o t e i n a t e i n representative types of s o i l s * another aspect of the r e l a t i o n between s o i l s and the f e r t i l i z e r i s evident i n the r e s u l t s of the l e a c h i n g and growing t e s t s . T h e o r e t i c a l l y , the r e t e n t i o n f a c t o r as d e r i v e d from growth response should be i n v e r s e l y p r o p o r t i o n a l to the degree of l e a c h i n g * i f the retained f e r t i l i z e r i s used with the same degree of e f f i c i e n c y In a l l s o i l s . That t h i s i s not the case i s shown by a comparison of the l e a c h i n g and r e t e n t i o n faotorst - 29 Leaching, Retenti on SOIL A (Ladner Clay) 2 7 SOIL B (Caster Loam) 10 4 SOIL 0 (Black Muck) 9 SOIL D (Upland Sandy Loam) 35 2 In the case of SOIL D l e a c h i n g was extremely high hut increase i n plant growth was a l s o extremely high. P o s s i b l e reasons f o r these r e s u l t s were s o i l f a c t o r s such as a c i d i t y , a e r a t i o n , and b i o l o g i c a l a c t i v i t y , a l l of which affected the rate of breakdown of the f i s h p r o t e i n a t e and i t s a v a i l a b i l i t y for p l a n t growth. In t h i s case the f e r t i l i s e r m a t e r i a l r e -tained i n the s o i l was used e f f i c i e n t l y and good plant growth r e s u l t e d . The opposite i s true i n the case of the heavier s o i l s ( SOILS A and B) from which l e a c h i n g was r e l a t i v e l y low, but increase i n plant growth was not h i g h . . Thus i t appears that p l a n t response to the a d d i t i o n of f i s h proteinate to the s o i l depends to a l a r g e degree upon the rate at which the nitrogenous m a t e r i a l i s made a v a i l a b l e to the p l a n t s , and a v a i l a b i l i t y i n turn depends upon the p h y s i c a l structure of the s o i l and i t s s u i t a b i l i t y f o r a c t i v e b a c t e r i a l decomposition of the p r o t e i n a t e . 5. C ONRUSH QKS, The r e s u l t s of t h i s experiment Indicate that the f i s h proteinate possesses some very d e s i r a b l e p r o p e r t i e s as a p o t e n t i a l nitrogen f e r t i l i z e r . In s p i t e of severe l e a c h i n g , - 30 - > i t i s strongly r e t a i n e d i n s o i l s , p a r t i c u l a r l y those of f i n e t e x t u r e . Although the rate of a v a i l a b i l i t y tends to be slow, e s p e c i a l l y i n heavy s o i l s , t h i s feature may be advantageous i n the case of c e r t a i n perennial or long-season crops f o r which an Immediate supply of nitrogen i s not e s s e n t i a l . This material should be s a t i s f a c t o r y as a source of nitrogen f o r l i g h t s o i l s when a p p l i e d on the surface because i t seems probable that the f e r t i l i z e r w i l l hot be leached by moderate r a i n f a l l beyond the depth of root growth, and that the rate of a v a i l a b i l i t y w i l l be s u f f i c i e n t l y high to provide adequately for sustained plant growth. In heavy s o i l s , where the nitrogen i s s t r o n g l y r e t a i n e d and only slowly a v a i l a b l e , i t seems that greater benefit would be derived If the material were incorporated i n the s o i l to some depth, rather than allowed to remain on the s u r f a c e . Although the f i s h p r o t e i n a t e has undeniable value as a source of nitrogen f o r plant growth, i t s use as a f e r t i l i z e r cannot be recommended at the present time. The tendency of the m a t e r i a l to absorb moisture from the a i r presents a serious d i f f i c u l t y . Even i f the m a t e r i a l could be maintained i n powder form by s p e c i a l care i n packaging and storage, the problem of a p p l i c a t i o n would remain, and i t i s evident from the l i m i t e d observations made during t h i s experiment that the f i s h proteinate could not be applied by ordinary mechanical means. A second drawback of the material is I t s tendency to form a hard, almost impervious c r u s t , p a r t i c u l a r l y on sandy s o i l . - 31 -It i s concluded, t h e r e f o r e , that u n t i l these undesirable p r o p e r t i e s of the material have been overcome, the f i s h proteinate should not be put on the market as a commercial f e r t i l i z e r . 6. SUMMARY i Water i n three separate a p p l i c a t i o n s was allowed to percolate through oolumns of pots c o n t a i n i n g d i f f e r e n t types of s o i l s to which f i s h proteinate had been added only i n the top p o t . The volume of water was s u f f i c i e n t to saturate a l l the s o i l and to give a small excess for a n a l y s i s . P l a n t s were subsequently grown i n a l l pots and the increased growth i n d i f f e r e n t pot l e v e l s was used as a measure of nitrogen r e t e n t i o n . On the whole the f i s h p r o t e i n a t e was well r e t a i n e d , p a r t i c u l a r l y i n heavy s o i l s , but the r e t a i n e d f e r t i l i z e r was more e f f i c i e n t l y u t i l i z e d i n l i g h t s o i l . The tendency of the f i s h p r o t e i n a t e i n i t s present form to absorb moisture from the a i r and to cause the formation of a hard c r u s t , e s p e c i a l l y i n the l i g h t s o i l , are d e f i n i t e draw-backs, and i t i s concluded that these undesirable features must be overcome before the product should be put on the market. - 32 -7. REFERENCES 1. Bray, R. H. N i t r a t e t e s t s for s o i l and p l a n t t i s s u e s . S o i l Science £0_» 123, 1945. 2. Conrad, J . P . Retention by the s o i l s of the nitrogen of various compounds as shown by subsequent plant growth. Journal of A g r i c u l t u r a l Research £0_s 617, 1940. 3. Bmmert, E . Iff. The r a p i d determination of t o t a l ni trogen i n s o i l . S o i l Science 58_s 289, 1944. 4. E e l l e y , C . C. and S p i l s b u r y , R. H. S o i l survey of the Lower F r a s e r V a l l e y . Canadat Department of A g r i c u l t u r e Technical B u l l e t i n 20, 1939. 5. T e l r , J . B. An evaluation of the f e r t i l i z i n g p r o p e r t i e s of a f i s h waste p r o d u c t . M a s t e r ' s T h e s i s , Department of H o r t i c u l t u r e , The U n i v e r s i t y of B r i t i s h Columbia, 1947. 6. Young, V. M. The determination of the value of a f i s h product f e r t i l i z e r . Unpublished Report, Department of H o r t i c u l t u r e , The U n i v e r s i t y of B r i t i s h Columbia, 1945. 

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