UBC Theses and Dissertations

UBC Theses Logo

UBC Theses and Dissertations

Fish waste in British Columbia and its conservation as fertilizer Merryfield, Jack William 1947

Your browser doesn't seem to have a PDF viewer, please download the PDF to view this item.

Item Metadata

Download

Media
831-UBC_1947_A4 M3 F5.pdf [ 9.23MB ]
Metadata
JSON: 831-1.0106964.json
JSON-LD: 831-1.0106964-ld.json
RDF/XML (Pretty): 831-1.0106964-rdf.xml
RDF/JSON: 831-1.0106964-rdf.json
Turtle: 831-1.0106964-turtle.txt
N-Triples: 831-1.0106964-rdf-ntriples.txt
Original Record: 831-1.0106964-source.json
Full Text
831-1.0106964-fulltext.txt
Citation
831-1.0106964.ris

Full Text

FISH WASTE IN BRITISH COLUMBIA And ITS CONSERVATION AS FERTILIZER by Jack W i l l i a m M e r r y f i e l d And S t u a r t W i l f r e d Turner 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 < AGRONOMY 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.. A C K N O W L E D G E M E N T S The authors wish to express their sincere appreciation to Dr. V. C. Brink for his assistance in outlining the experimental work, his helpful criticism, and his review of this paper; to Dr. W. C. Chalmers for providing the problem and the material; to Miss M. 0. Burton for her microbiological report; to the Staff of the Department of Agronomy for their timely and helpful suggestions; to Mr. F. D. Cook for the use of the digest agitator; to Dr. A. M. Crooker for carrying out the spectographic analysis; and to Mr. M. Clarke, Mr. I . J . McDonald, Mr. G. Reifel, Mr. D. Swackhammer, and the Seal-Sap Farms for their cooperation i n the field t r i a l s . - 0 O 0 C O N T E N T S I n t r o d u c t i o n : •—Page I M a t e r i a l Page 4 Problems Undertaken - F e r t i l i z e r Value —Page 5 A Comparison of P r o t e i n a t e pH 12, 10, 8.5, and 6 w i t h and without P2O5 and K2O Supplements w i t h Commercial Organic & Inorganic F e r t i l i z e r s Page 7 Greenhouse T r i a l — Page . 7 F i e l d T r i a l s Page 20 Oat Test '• Page 20 Sugar Beet Seed Test Page 25 Pasture Test Page 27 Lawn Test Page 29 A Study of the E f f e c t of P r o t e i n a t e of S e v e r a l pHs on the Germination, S e e d l i n g Development, and Root Formation of s e v e r a l P l a n t Species Page 30 A Study of the Absorption & Movement of P r o t e i n a t e i n t h i s s o i l & i t s r e s u l t a n t E f f e c t on the Vegetation Page 31 A Report on P r o t e i n a t e F e r t i l i z e r i n R e l a t i o n to B a c t e r i a l Growth Page 32 A Comparison of the E f f e c t of Leaching on' P r o t e i n a t e w i t h the E f f e c t on Commercial F e r t i l i z e r • Page 43 Laboratory T r i a l Page 43 F i e l d T r i a l • Page 44 A Determination of the Minor Element Content of P r o t e i n a t e Page 47 ' An I n v e s t i g a t i o n as to the S u i t a b i l i t y of P r o t e i n a t e t o Pass Through the Standard F e r t i l i z e r . A p p l i c a t -ions Page 49 I n v e s t i g a t i o n i n t o the Problem of P r o t e i n a t e Deliquescence page 50 C O N T E N T S - (Continued) A Method of Preparing a Complete N.P.K. F e r t i l i z e r with the Proteinate Base. —Page 52 Problems Undertaken - Feeding Value Page 6 The Determination of the Protein Index of the Proteinate — 1 —;—Page 54 A Study of the Effect of Proteinate on the Palatability of Various Fodder Crops Page 59 Discussion Page 60 Conclusion Page 62 Literature Cited Page 63 ILLUSTRATIONS I l l u s t r a t i o n (1) - Greenhouse T r i a l (January 26, 1946) I l l u s t r a t i o n (3) - Greenhouse T r i a l (February 21, 1946) I l l u s t r a t i o n (3) - Upland S o i l R e s u l t s I l l u s t r a t i o n (4) - Lowland S o i l R e s u l t s I l l u s t r a t i o n (5) - I n t e r i o r S o i l R e s u l t s I l l u s t r a t i o n (6) - Oat Test P l o t I l l u s t r a t i o n (7) - H a r v e s t i n g Sugar Beet Seed Test I l l u s t r a t i o n (8) - Apparatus f o r Determination o f Undigestable P r o t e i n . TABLES Table (1) - Greenhouse T r i a l Dry Weights Table (2) - S t a t i s t i c a l R e s u l t s on the Upland S o i l Table (3) - S t a t i s t i c a l R e s u l t s on the Lowland S o i l Table (4) - S t a t i s t i c a l R e s u l t s on the I n t e r i o r S o i l Table (5) - Oat Test Y i e l d Table (6) - S t a t i s t i c a l R e s u l t s o f the Oat Test Table (7) - Sugar Beet Seed Y i e l d Table (8) - S t a t i s t i c a l R e s u l t s of Sugar Beet Seed Test Table (9) - B a c t e r i a Growth measurement i n 3 days Table (10) - B a c t e r i a Growth measurement i n 5 days Table (11) - B a c t e r i a Growth measurement i n 14 days Table (12) - F i e l d Leaching T r i a l R e s u l t s . ABSTRACT An i n v e s t i g a t i o n has been c a r r i e d out on the value and use of f i s h waste as a f e r t i l i z e r . The m a t e r i a l i s a p r o t e i n a t e c o n t a i n i n g 10% n i t r o g e n and o r i g i n a t e s as " o f f a l " , a by-product of the v i t a m i n o i l e x t r a c t i o n process. The p r o t e i n a t e has been compared w i t h commercial organic and i n -organic f e r t i l i z e r s and c e r t a i n conclusions have been reached. In a d d i t i o n , the minor element content, the manu-fa c t u r e of a complete f e r t i l i z e r , and the p o s s i b l e f e e d i n g value of the m a t e r i a l have been i n v e s t i g a t e d and c e r t a i n conclusions drawn. INTRODUCTION For years, time and thought has been given t o the u t i l i z a t i o n o f f i s h wastes e i t h e r as f e r t i l i z e r s , feed supp-lements, or v i t a m i n o i l s . I n B r i t i s h Columbia, e s p e c i a l l y , t e c h n i c a l personnel have been keenly aware o f t h i s g r eater development, p r i m a r i l y because of the importance of the F i s h i n g I n d u s t r y i n t h e i r p r o v i n c e , and secondly because o f the l a r g e q u a n t i t i e s o f by-products which as yet have not been f u l l y u t i l i z e d . At the present time the extreme shortage of organic m a t e r i a l s f o r f e r t i l i z a t i o n programs coupled w i t h the world's demand f o r i n c r e a s e d food p r o d u c t i o n , compels a thorough i n v e s t i g a t i o n and immediate development o f a l l p o s s i -b l e sources of p l a n t n u t r i e n t s . Increased bushels per acre and increased acreages o f food crops are both e s s e n t i a l i n meeting t h i s emergency. The m a t e r i a l s t u d i e d i n t h i s -paper may be one answer t o our organic f e r t i l i z e r needs. I t i s a sodium p r o t e i n a t e r e s i d u e from one o f the f i s h o i l e x t r a c t i o n processes o f B r i t i s h Columbia's F i s h i n g I n d u s t r y . Many r e p o r t s have been published on the use o f f i s h « wastes as f e r t i l i z e r s , (4) (5) (7) ( 8 ) , and c e r t a i n i n v e s t i g a -t i o n s have been c a r r i e d out t o determine i t s value as a feed supplement, (1) (2) (6>). Other workers have s t u d i e d the chemistry and technology o f marine animal o i l s (3) i n an attempt to f u r t h e r the i n f o r m a t i o n on f i s h wastes, as t o t h e i r - 2 -value and uses. However, the f i e l d of study and experimentat-i o n i s f a r from exhausted. The o l d e s t i n d u s t r y to make use of waste f o r f e r t i l -i z i n g purposes i s the f i s h i n d u s t r y (7 ) . The custom of f e r t i l -i z i n g crops w i t h f i s h e x i s t e d among the Indians of New England l o n g before the a r r i v a l of the white s e t t l e r s i n North America. I t i s s a i d t h a t r o r f e r t i l i z i n g corn, one o r two f i s h were placed i n each h i l l . This procedure was adopted by the c o l o n i s t s , and a t the time o f a l a r g e c a t c h the s u r p l u s f i s h were simply spread broadcast over the f i e l d s . I n p l a c e s where f i s h s u p p l i e s have been i n abundance, the d i s p o s a l of the oversupply has been p r a c t i c e d as such u n t i l recent times. I t was soon observed however, that the response f i r s t noted on l i b e r a l l y f e r t i l i z i n g w i t h r i s h decreased w i t h each successive a p p l i c a t i o n and the i n i t i a l good e f f e c t s could be obtained again o n l y a f t e r the s o i l had been allowed t o stand f o r a time without f u r t h e r a p p l i c a t i o n of the f i s h . A f t e r many suggest-ions had been advanced.as an e x p l a n a t i o n of t h i s r e s u l t , i t was f i n a l l y shown t h a t the d e l e t e r i o u s a c t i o n on the p l a n t s was owing t o the o i l i n the f i s h . When the o i l was removed by cooking and p r e s s i n g , the residue obtained no longer e x h i b i t e d the e f f e c t s p r e v i o u s l y noted, and i t s value as a f e r t i l i z e r was consequently g r e a t l y improved. A f u r t h e r advantage i n removing the o i l was seen i n the p r o p e r t i e s which the o i l possesses i n i t s e l f f o r use i n i n d u s t r y and v i t a m i n products. I t was vsoon r e a l i z e d t h a t the returns from crude f i s h o i l o f f i s h wastes were more than s u f f i c i e n t to cover the cost of e x t r a c t i o n . At present, the recovery of the residue o f the e x t r a c t i o n process o f t e n becomes subordinate to the recovery of the o i l . The purpose of t h i s paper i s t o determine the value and p o s s i b l e use of a c e r t a i n residue from the f i s h o i l e x t r a c t i o n processes i n B r i t i s h Columbia now being c a r r i e d out. - 4 -MATERIAL The m a t e r i a l under c o n s i d e r a t i o n o r i g i n a t e s as the *Offal» obtained as a by-product from the f i s h canning indus-t r y . At one time, and t o a l e s s e r extent a t the present time, most o f the wastes from the canneries were merely dumped i n t o the ocean. Of l a t e a c e r t a i n p o r t i o n of t h i s waste has been d i r e c t e d i n t o other channels and has been found t o be of great v a l u e . By means o f an e f f i c i e n t process, the v i t a m i n c a r r y i n g o i l s are e x t r a c t e d from the o f f a l and an a l k a l i n e m a t e r i a l , a sodium p r o t e i n a t e r a t h e r h i g h i n n i t r o g e n , remains as a r e s i d u e . The o i l has been the prime o b j e c t i v e i n the process and as a consequence the r e s i d u e has been discarded and considered as useless, owing t o i t s undecided value as a f e r t i l i z e r and the p o s s i b l e d i f f i c u l t i e s of pr o c e s s i n g i t as such. By means of experimental t e s t s , the va l u e o f t h i s r e s i d u e as a f e r t i l i z e r w i l l be determined. The m a t e r i a l c o n s i s t s of p r o t e i n s i n v a r i o u s stages of degradation. I t i s a dark Drown sludge w i t h a strong odor of decomposed f i s h , s o l i d a t low temperatures (20 degrees C.) and a v i s c o u s l i q u i d a t h i g h temperature (37 degrees C ) . The question a r i s e s - would i t be economical o r f e a s i b l e to handle the m a t e r i a l i n the l i q u i d form or should i t be t o t a l l y con-v e r t e d t o the conventional s o l i d form? The o i l e x t r a c t i o n • process e f f e c t s the l o s s of hydroxyl groups w i t h the r e s u l t t h a t the residue tends t o be ve r y d e l i q u e s c e n t . This i n i t s e l f presents q u i t e a problem i f the m a t e r i a l i s ever t o be o f use as a f e r t i l i z e r . Another f e a t u r e i s that i t gives a r e a c t i o n of pH 12. Perhaps i t would he advisable to reduce the a l k a l i n i t y but to what pH? Then again should s u l p h u r i c a c i d , which i s r e l a t i v e l y cheap but which would form sulphates t o the extent of 30%, or phosphoric a c i d , which i s r e l a t i v e l y expensive, but which would form phosphates thus i n c r e a s i n g the f e r t i l i z e r v a l u e , be used i n the process of n e u t r a l i z a t i o n ? PROBLEMS UNDERTAKEN The f o l l o w i n g problems have been s t u d i e d i n order t o appraise the value of p r o t e i n a t e as a f e r t i l i z e r . The p r o t e i n a t e was compared w i t h commercial organic and in o r g a n i c f e r t i l i z e r s towards t h i s end. Other f e a t u r e s such as e f f e c t s on s o i l microorganisms, s u i t a b i l i t y f o r storage, ease of app-l i c a t i o n , development of a process f o r the manufacture of a complete N.P.K. f e r t i l i z e r , and the e f f e c t of the p r o t e i n a t e on v a r i o u s s o i l s , were a l s o considered. FERTILIZER VALUE 1. A comparison o f p r o t e i n a t e pH 12, 10, 8 . 5 , and 6, w i t h and without P2O5 & K2O supplements, w i t h commercial organic and in o r g a n i c f e r t i l i z e r s . 2. A study of the e f f e c t of p r o t e i n a t e , a l l mentioned pHs, on the germination, s e e d l i n g development, and root formation of s e v e r a l p l a n t s p e c i e s . 3. A study of the absorption and movement of p r o t e i n a t e i n the s o i l and i t s e f f e c t s on the v e g e t a t i o n . 4. A study o f the e f f e c t of p r o t e i n a t e on the s o i l microorganisms. 5 . A comparison o f the e f f e c t of l e a c h i n g p r o t e i n a t e t r e a t e d s o i l , w i t h the e f f e c t of l e a c h i n g commercial f e r t i l i z e r t r e a t e d s o i l . 6. A determination of the minor element content o f p r o t e i n a t e . 7. An i n v e s t i g a t i o n of the s u i t a b i l i t y of p r o t e i n a t e to pass through the standard f e r t i l i z e r a p p l i c a t o r s , 8. I n v e s t i g a t i o n s i n t o the deliquescence of p r o t e i n a t e . FEEDING VALUE • ' . 1. Determination of the p r o t e i n index of p r o t e i n a t e . 2. A study of the e f f e c t of p r o t e i n a t e on p a l a t a b i l i t y o f v a r i o u s fodder crops. To d i s c u s s f i s h by-products from any p o i n t o f 1 v i e w other than t h a t of f e r t i l i z e r s , i s perhaps beyond the scope of t h i s paper. However, the p r o t e i n a t e has shown promise as a feed supplement and as such i s worthy of some a t t e n t i o n . The l i m i t e d t e s t s along t h i s l i n e were as l i s t e d above. - 7 A COMPARISON OF PROTEINATE pH 18, 10, 8*5, and 6, WITH * • AND WITHOUT P2O5 & Kafr SUPPLEMENTS, WITH COMMERCIAL  ORGANIC AND INORGANIC FERTILIZERS The primary c o n s i d e r a t i o n i n the e v a l u a t i o n o f pro-t e i n a t e as a f e r t i l i z e r was "how does i t compare w i t h commer-c i a l f e r t i l i z e r s " . This comparative study was handled i n two phases. A greenhouse t e s t under c o n t r o l l e d l i g h t and tempera-ture c o n d i t i o n s and as an immediate i n d i c a t i o n of the r e l a t i v e value of the p r o t e i n a t e t o organic and inor g a n i c f e r t i l i z e r s was f i r s t e s t a b l i s h e d . Secondly, a number of f i e l d t e s t s were made. These were made t o study the e f f e c t s o f the p r o t e i n a t e on seed p r o d u c t i o n , pasture development, and lawn growth. GREENHOUSE TRIAL' Samples of p r o t e i n a t e : -A. Unen^tralized m a t e r i a l pH 12 B. N e u t r a l i z e d and Diatomaceous e a r t h pH 10 C. " " " " pH 8.5 D. " tt " " pH 6 Combinations and r a t e s of a p p l i c a t i o n : -1. 2. 4 samples o n l y - 8$ N 4 samples & P205~8% N 750#/A -750#/A -60#N/A 60#N/A 4. 3. 4 samples & P2O5 & K 20 -8% N 4 samples & Ca -8% N 750#/A -750#/A -858#/A -300#/A -60#N/A 60#N/A 60#N/A 60#N/A 5. Blood & Bone -7% N 6. (NH4) 2S04 20% N I - 8 -7. M i l l o r g a n i t e - 6% N — 1000#/A — 8. N.P.K. (8-10-5)- 8% N — 750#/A — 600/A 9. C o n t r o l - N I L NB. Rate of a p p l i c a t i o n was based on an acre f o o t weight of mi n e r a l s o i l - 2,000,000 l b s . The P2O5 & KgO supplements were added i n equi v a l e n t s to the N.P.K. mix. The Ca supplement was added at the r a t e of 500#/A. ffoils used:-1. I n t e r i o r ( a l k a l i ) s o i l (Summerland Area) 2. Lowland s o i l ( F r a s e r V a l l e y D e l t a Area) 3. Upland S o i l (U.B.C. Area) T e s t i n g crop:-O l l i B a r l e y No. 140 - 1943, t r e a t e d f o r smut & s t r i p e c o n t r o l . Design:- Randomized block, 3 R e p l i c a t i o n s . P o t s i z e : - 9£" x 5" (To hold 4" depth of s o i l ) No. o f p l a n t s per pot:- 20 (Seed 25 - t h i n t o £0) Bench Dimensions:- 5*8" x 20*10" (This allows pot t e s t to be set up 7 by 27 pots as randomized 3 r e p l i c a t i o n d e s i g n ) . Treatments:-1. Sample A 2. » B 3. » C 4. " D 5. " A & P 2 0 5 - 9 6. Sample B & *2°5 7. C & P g 0 5 8. « D & P 2 O 5 9. n A & P 2 0 5 & KgO 10. n B & P g 0 5 & KgO 11. H C & P g 0 5 & KgO 12. tt D & P g 0 5 & KgO 13. tt A & Ca 14. tt B & Ca 15. tt C & Ca 16. tt D & Ca 17. M i l l o r g a n i t e 18. Blood and Bone 19. ( N H 4 ) 2 S 0 4 20. N.P.K. mix 21. C o n t r o l • M o i s t u r e , Temperature and L i g h t ; -M o i s ture and temperature were c o n t r o l l e d zo promote optimum growth. A u x i l i a r y l i g h t i n g of two 2000 watt G.'E. f l u o r e s c e n t lamps were suppli e d t o a i d i n m a i n t a i n i n g a favourable photosynthesis - r e s p i r a t i o n balance. Procedure:-On January 26th, 1946, the s o i l s were mixed and screened and 8# placed i n pot. The treatment.materials were incorporated i n the s o i l and the pots were placed on the beneh i n the greenhouse according t o the randomized d e s i g n . - 10 -On February 8 t h , the B a r l e y was seeded at the ra t e of 25 seeds per p o t . The seedlings were thinned t o 20 p l a n t s per pot on February 21st. Observations on height and c o l o u r were made b i -monthly from March 5th t o A p r i l 5th when the crop was harvested and .the dry weights o f the tops were recorded. A s t a t i s t i c a l a n a l y s i s o f the r e s u l t s was c a r r i e d out. - 11 -GREENHOUSE TRIAL - January 26, 1946  ILLUSTRATION (1) GREENHOUSE TRIAL - February 21, 1946  ILLUSTRATION (2) TABLE I - 15 -GREENHOUSE TEST - Dry weights of Tops from A l l Treatments and Blocks (gms.) e 1 Treat. 1 Totals | CM CO o> t> 03 CO LO co 12.7 12.8 13.3 13.0 11.8 13.2 15.0 13.1 to CO CO 17.8 | 03 0> in CO cO CO H 5D co to rH 03 OO 216.1 1 Rows • 1 H H H O 03 m O CO to CO o CO in H cO co H o 03 cO in o 03 CO 03 in 03 co H CO in co H 03 CO X! oo 03 o CV3 O 1 Rows • 1 H H 03 CO o CO in 03 co CV3 o in o in 03 o CO cn 03 03 CO o to o> 03 H CO o CO o> 03 CO A3 O in > 03 • to o 1 Rows • 1 H CO 03 03 t> 03 to 03 t> to CO O m o o CO in «tf 03 tO 03 03 o CV3 03 03 S3 O-H in in O 03 70.0 LOWLAND 1 Treat.1 1 Totals CN O o CO a. O O H CO CO 10.5 12.1 03 cn 10.3 10.6 10.3 10.7 11.5 | CO O H 12.3 I LO H H 0* 0> (O CO CO A3 H in > 213.1 LOWLAND 1 Rows H H H co 03 CO 03 in co in co in CO o to o CO co H CO o CO H to m CO cn to cO co 03 03 CO A3 CO O to CO 03 72.0 LOWLAND 1 Rows HH H o> 03 in 03 o> 03 o CO CO 03 CO CV2 LO CO CO co CO CO co CO 03 CO 03 CO o ^» o CO in CO to CO K 03 H in co 03 H o LOWLAND 1 Rows H in CO CO o CO m CO rH CO O co o co o co 03 o CO CO o to 03 to cO to CO CO CO co CO to CO t> 03 o cn 03 71.0 INTERIOR 1 Treat. 1 Totals 14.6 16.2 o o H 15.5 03 O i—1 o 03 18.5 15.7 CO m H 03 o> H CO o H 18.3 03 CO rH o-m H 03 cO H 16.7 | cO H CO to H H <tf» 02 26.9 J O 03 H 369.4 INTERIOR | Rows H H H 03 in o in m CO o o O «o t> in o to o in cO CO in H O O in CS LO CO in OO in m in o-H cn rH 117.9 INTERIOR | Rows H H o» • o • in o CO • LO H • m CO o i—i • cO cO in CO in cO 03 . to co cO H • in in in • m CO • m to • in CO . in co CO CO o • 121.1 INTERIOR | Rows H co in o (0 CO o 03 co H in CO O cO CO cO in CO CO CO cO tO tO o to CO o in H in to tO m in CO 0> o o H in co • o CO o 9 * H O cc r™ OC +: C E-Treat-ments 1 03 CO in CO O CO cn o H H H 03 H to H H in H cO H o H CO H cn H O P3, H 03 - 16 -STATISTICAL RESULTS ON THE UPLAND SOIL TABLE I I Source o f V a r i a t i o n Sum of Squares Degrees of Freedom Variance F Value 5^ P t . of F i i p t of I Treatments 46.3193 20 2.3159 4.8087 1.83 2.37 Blocks .3069 2 .1534 .3185 3.23 5.17 E r r o r 19.2665 40 .4816 T o t a l V a r i a -b i l i t y 65.8927 62 Since the " F w value of treatments i s l a r g e r than the 5% p o i n t of n F " , s i g n i f i c a n t treatment d i f f e r e n c e s are present. Treatment Means: 1. 2.73 8. 4.33 15. 2.6 2. 2.63 9. 3.93 16. 3.06 3. 2.73 10. 4.4 17. 2.83 4. 2.83 11. 5.0 18. 2.86 5. 4.23 12. 4.37 19. 2.03 6. 4.26 13. 2.76 20. 4.43 7. 4.76 14. 2.8 21. 2.73 Minimum d i f f e r e n c e r e q u i r e d f o r s i g n i f i c a n c e : / .9652 x 1.9599 = 1.1104 V 5 CONCLUSIONS: 1. A l l treatments, w i t h the exception o f 1, 2, 5, 4, 13, 14, 15, 16, 17, 18, 19, o u t y i e l d the c o n t r o l s i g n i f i -c a n t l y . 2. No s i g n i f i c a n t d i f f e r e n c e s e x i s t between p r o t e i n a t e pHs. - 17 -3. No s i g n i f i c a n t d i f f e r e n c e s e x i s t between p r o t e i n a t e and Other o r g a n i c s . 4. No s i g n i f i c a n t d i f f e r e n c e s e x i s t between p r o t e i n a t e and ( N H 4 ) g S 0 4 . 5. P r o t e i n a t e & P2O5 showed increased y i e l d s compared w i t h p r o t e i n a t e . 6. P r o t e i n a t e & P2O5 & K2O showed increased y i e l d s compared w i t h p r o t e i n a t e . 7. No s i g n i f i c a n t d i f f e r e n c e s e x i s t between p r o t e i n a t e p l u s Ca and p r o t e i n a t e . 8. N.P.K. (8-10-5) r e s u l t e d i n s i g n i f i c a n t y i e l d increases over a l l treatments except 5 to 12 i n c l u s i v e . STATISTICAL RESULTS ON THE LOWLAND SOIL  TABLE I I I Source of V a r i a t i o n Sum of Squares Degrees of Freedom Variance F Value 5% P t . of F 1* P t of F Treatments 13.9041 20 .5695 3.88 1.83 2.37 Blocks .0860 2 .0430 .29 3.23 5.17 E r r o r 5.8607 40 .1465 T o t a l Varia-b i l i t y 19.8508 62 v Since the "F" value o f treatments i s l a r g e r than the 5f?o p o i n t o f rtF", s i g n i f i c a n t treatment d i f f e r e n c e s are present. Treatment Means: 1. 2.56 4. 3.56 7 . 3.5 2. 2.9 5. 3.13 8. 4.03 3. 3.13 6. 2.93 9 . 3.06 10. 3.63 14. 3.83 18. 3.3 11. 3.53 15. 3.53 19. 2.86 12. 3.43 16. 4.1 20. 4.26 13. 3.56 17. 3.83 21. 2.5 Minimum d i f f e r e n c e r e q u i r e d f o r s i g n i f i c a n c e : 2930 x 1.9599 .612 CONCLUSIONS: 1. A l l treatments w i t h the exception of 1, 2, 9, and 19, o u t y i e l d the c o n t r o l s i g n i f i c a n t l y . 2. pH 6 showed increased y i e l d compared w i t h p r o t e i n a t e pH 10 and pH 12. 3. M i l l o r g a n i t e and Blood & Bone o u t y i e l d p r o t e i n a t e pH 12. M i l l o r g a n i t e a l s o o u t y i e l d s p r o t e i n a t e pH 10. 4. P r o t e i n a t e pH 6 o u t y i e l d s (NILiJgSO^ 5. No s i g n i f i c a n t d i f f e r e n c e s e x i s t between p r o t e i n a t e p l u s P2°5 and p r o t e i n a t e a l l pHs. 7. P r o t e i n a t e pH 12 p l u s Ca o u t y i e l d p r o t e i n a t e pH 12. P r o t e i n a t e pH 10 p l u s Ca o u t y i e l d p r o t e i n a t e pH 10. 8. N.P.K. (8-10-5) showed no s i g n i f i c a n t y i e l d d i f f e r e n c e over p r o t e i n a t e pH 6 p l u s P2O5, p r o t e i n a t e pH 10 p l u s Ca, p r o t e i n a t e pH 6 p l u s Ca, or M i l l o r g a n i t e . STATISTICAL RESULTS ON THE INTERIOR SOIL TABLE IV (See next page) 6. P r o t e i n a t e pH 10 p l u s P2O5 p l u s KgO o u t y i e l d p r o t e i n a t e pH 10. - 19 -Source of V a r i a t i o n Sum o f Squares Degrees of Freedom Variance F Value 5% P t . of F. 1% P t o f 5 Treatments 68.338 20 3.4169 5.135 1.83 2.37 Blocks 4.0145 2 2.0072 3.017 3.23 5.17 E r r o r 26.6125 40 .6653 T o t a l Varia-' b i l i t y 98.965 62 Since the " F n Value of treatments i s . l a r g e r than the 5% p o i n t of M F " , the d i f f e r e n c e i s s i g n i f i c a n t . The blocks are a l s o s i g n i f i c a n t . Treatment Means: 1. 4.86 7. 6.16 13. 6.06 19. 8.03 2. 5.4 8. 5.23 14. 5.23 20. 8.96 3. 5.9 9. 5.20 15. 5.4 21. 4.0 4. 5.16 10. 6.4 16. 5.56 5. 5.73 11. 5.76 17. 5.46 6. 6.86 12. 6.1 18. 5.6 Minimum d i f f e r e n c e r e q u i r e d f o r s i g n i f i c a n c e : V 1.3506 X 1.9599 = 1.307 3 CONCLUSIONS: 1. A l l treatments w i t h the exception of 1,4,8,9, and 14 o u t y i e l d c o n t r o l . 2. No s i g n i f i c a n t y i e l d d i f f e r e n c e s e x i s t between p r o t e i n -ate pHs. 3. No s i g n i f i c a n t y i e l d d i f f e r e n c e s e x i s t between p r o t e i n -ate and other organic f e r t i l i z e r s . 4. (NH 4)gS0 4 o u t y i e l d p r o t e i n a t e . - 20 -5. P r o t e i n a t e pH 10 p l u s Pg Oj o u t y i e l d p r o t e i n a t e pH 10. 6. No s i g n i f i c a n t y i e l d d i f f e r e n c e e x i s t s between p r o t e i n -ate p l u s P2O5 p l u s KgO and p r o t e i n a t e . 7. No s i g n i f i c a n c e . 8. N.P.K. (8-10-5) o u t y i e l d a l l p r o t e i n a t e treatments. GENERAL CONCLUSION As may be noted i n the f o r e g o i n g c o n c l u s i o n s , the p r o t e i n -ate exerted d i f f e r e n c e e f f e c t s on the three s o i l t y p e s . This i s i n keeping w i t h most f e r t i l i z i n g m a t e r i a l s . P r o t e i n a t e s u p p l i e s a N i t r o g e n source f o r p l a n t growth and i n t h i s respect compares w e l l w i t h organic f e r t i l i z e r s . No d e l e t e r -ious e f f e c t s on p l a n t growth were e x h i b i t e d by the p r o t e i n a t e . Thus, from the greenhouse t e s t we may conclude t h a t p r o t e i n a t e has a decided value as a f e r t i l i z e r as compared to commercial f e r t i l i z e r s . OAT TEST: Treatments: 1. C o n t r o l 2. P r o t e i n a t e pH 8.5 500#/A ...40#N/A 3. P r o t e i n a t e pH 8.5 & P 2 0 5 & K 20 ...650#/A ...40#N/A 50#P205/A 25#K2 0/A 4. M i l l o r g a n i t e 675#/A ...40#N/A 5. N.P.K. (8^10-5) mix 500#/A .. .40#N/A 50#P205/A 25#K 20/A - 21 -6. (NH4) 2S0 4 200ft/k ... .4Q#N/A L o c a t i o n of p l o t s and s o i l types; 1. A g a s s i z - mlsed. c l a y p r o f i l e . 2. Crescent Beach - Ladner c l a y w i t h some peat. 3. Mount Lehman - Whatcom sandy loam. T e s t i n g crop: - V i c t o r y Oats No. 5 Design: Randomized block, 3 R e p l i c a t i o n s . S i z e o f p l o t s : - 5' x 5» (25 sq.. f t . ) Rate of seeding:- 1.5 bu. - A c r e . Procedure:- The p l o t s were prepared on ground t h a t had not been f e r t i l i z e d r e c e n t l y , p r e f e r a b l y not w i t h i n the past f o u r to f i v e y e a r s . The s e l e c t e d s i t e s were ploughed, t h o r -oughly d i s c e d , and marked o f f i n t o treatment rows w i t h stakes. Seeding and treatment a p p l i c a t i o n s were c a r r i e d out as f o l l o w s : A g a s s i z May 1 s t , 1946 Crescent Beach May 12th, 1946 Mount Lehman May 19th, 1946 Observations were made monthly w i t h p l a n t c o l o u r , h e i g h t , v i g o r , and presence of s t o o l i n g as the prime c o n s i d e r a t i o n s . The harvest dates were as f o l l o w s : A gassiz August 18th, 1946 (maturity) Crescent Beach • August 21st, 1946 ( " ) Mount Lehman August 19th, 1946 ( " ) The oats were p l a c e d - i n stooks f o r two favourable days and then threshed. The y i e l d of oats-was then recorded and the r e s u l t s analyzed s t a t i s t i c a l l y . - 23 -TABLE V - Y i e l d of Oats from A l l l o c a t i o n s (cleaned g r a i n i n ounces) AGASSIZ Row 1 2 T R E 3 A T M E 4 N T S 5 J 6 Row T o t a l s I I I I I I 6.5 9.0 6.5 11.5 7.5 12.5 7\0 8.5 8.0 14.5 7.0 12.5 12.5 ] 15.5 10.0 10.0 15.5 16.5 62.0 63.0 66.0 Treat. T o t a l s 22.0 31.5 23.5 34.5 38.0 42.0 191.0 CRESCENT BEACH I I I I I I 11.5 8.5 12.5 11.0 10.0 16.0 12.5 14.5 13.5 17.0 15.0 13.5 16.5 14.0 15,. 5 15.5 11 Jo 1 18.0 84.0 73.0 89.0 Tr e a t . T o t a l s 32.5 37.0 40.5 45.5 46.0 j-44.5 1 246.0 MOUNT LEHMAN I I I I I I 9.5 7.0 L1.0 12.0 8.0 8.'0 16.0 13.0 12.0 9.0 12.5 13.0 8.5 14.5 17.0 12.5 9.5 18.0 67.5 64.5 79.0 T o t a l s 27.5 28.0 41.0 34.5 40.0 40.0 ' 211.0 T o t a l Treat ment TOTALS 82.0 96.5 105.0 114.0 124 .:0 126.5 648.0 T o t a l Treat ment MEANS 9.1 10.7 11.6 12.6 13.7 • > 14.05 - 24 -STATISTICAL RESULTS OF THE OAT TEST TABLE VI Source of V a r i a t i o n Sum o f Squares Degrees o f Freedom Variance F-Value 5% P t . of F Treatments 161.277 5 32.255 4.072 2.71 Blocks 31^.694 2 15.847 2.413 . 4.10 Locations 86.111 2 43.055 Treatments X Blocks 65.669 10 6.566 Treatments X Locations 55.232 10 5.523 Block X * Locations 11.58 4 2.897 -Treatments X Blocks X Locations 158.437 20 7.921 T o t a l V a r i a b i l i t y 570.000 53 Since the "F" value f o r treatments i s l a r g e r than the 5% poin t of "F", the d i f f e r e n c e i s s i g n i f i c a n t . Treatment. Means: 1. 9.11 4. 12.66 2. 10.72 5. 13.77 3. 11.66 6. 14.05 Minimum d i f f e r e n c e r e q u i r e d f o r s i g n i f i c a n c e : /.15.842 x 2.086. s 2.768 V § CONCLUSIONS: 1. Treatments 4, 5, and 6 o u t y i e l d the c o n t r o l s i g n i f i -c a n t l y . - 25 -2. Treatments 5 and 6 o u t y i e l d treatment 2 s i g n i f i c a n t l y . 3. There were no s i g n i f i c a n t d i f f e r e n c e s between treatment No. 3 and a l l other treatments. 4. Treatments 2.and 3 showed no s i g n i f i c a n t y i e l d d i f f e r -ences over c o n t r o l . SUGAR BEET SEED TEST  Treatments Rate of A p p l i c a t i o n per Acre 1. C o n t r o l 2. P r o t e i n a t e pH 12 ( l i q u i d ) 40# 3. » . . » 80# 4. " " & P 20 5& KgO 40# 50# 25# 5. » . " » & " & " 80# 50# 25# L o c a t i o n of p l o t and s o i l type: Westham I s l a n d , Ladner, B.C. - Ladner C l a y T e s t i n g Crop: Sugar Beets i n 2nd year (seed production) Design: L a t i n Square S i z e o f P l o t s : - 20' x 20* - (100' x 100' T o t a l p l o t ) . Procedure:- The treatments were, l a i d out according to the p l a n on the 2nd year (seed year) Sugar Beets on May 18, 1946. Observations were made monthly u n t i l c u t t i n g on August 4 t h , 1946. The p l o t s were threshed on August 2 1 s t , 1946 and seed Y i e l d s recorded. - 26 -ILLUSTRATION 7 HARVESTING SUGAR BEET SEED SUGAR BEET SEED TEST PLOT DESIGN R 1 2 5 4 1 3 R 2 1 2 5 3 4 R 3 5 1 3 4 2 R 4 4 3 2 5 1 R 5 3 4 1 2 5 / - 27 -TABLE V I I - Y i e l d of Sugar Beet Seed (#/plot) R 1 Zlf 24 30 21 22 R 2 25 . 24 26 26 20 R 3 25 23 24 26 22 R 4 25 25 29 29 20 R 5 * 23 21 25 26 24 STATISTICAL RESULTS OF THE SUGAR BEET TEST TABLE V I I I Source of V a r i a t i o n Sum of Squares Degrees of Freedom Variance F Value 1 5% P t . o f " Treatment Row Column E r r o r ( u n c o n t r o l l 26.592 94.992 15.392 60.816 ed) 4 4 4 12 6.648 23.748 3.848 5.068 1.31 3.26 3.86 5.91 T o t a l 1 V a r i a b i l i t y 97.792 • 24 Since the treatment value of "F" i s l e s s than the 5$ poi n t of F, i t can be assumed th a t no s i g n i f i c a n t d i f f e r e n c e s e x i s t between treatments. Thus i t i s p o s s i b l e to conclude t h a t the p r o t e i n a t e pH 12, l i q u i d form, had no b e n e f i c i a l e f f e c t i n the P r o d u c t i o n o f Sugar Beet Seed. PASTURE TEST Treatments Rate of A p p l i c a t i o n per Acre H PiT5i *50 1. P r o t e i n a t e pH 6.5 (powder) 40# 2. » » " 80# 50# 50# 3. P r o t e i n a t e pH 6.5 (powder) 40# - 25# 4. ft tt tt so# : - 25# 5. tt tt tt 40# 50# 25# 6. tt tt tt 80# 50# 25# 7. tt pH 12 ( l i q u i d ) 40# 50# -8. tt tt H 80# 50# -9. tt tf tt 40# - 25# l O - ft tt ft 80# - 25# l l . tt i t tt 40# 50# 25# 12. tt tt tt 80# 50# 25# 13. C o n t r o l L o c a t i o n o f p l o t and s o i l type: S e a l Kap Farm - M i l n e r C l a y Loam. Te s t i n g crop: Red C l o v e r , White C l o v e r , 'Canada Bluegrass and P e r e n n i a l Ryegrass. Design:- Randomized S t r i p s . S i z e o f p l o t s : - 8' x 20' Method o f A p p l i c a t i o n : Broadcast Procedure:- The t r a i l was l a i d out according t o p l a n on May 24th, 1946. L i v e s t o c k were excluded from the area u n t i l August 6th, 1946. Observations were made monthly from May 24th, 1946 to September 12th, 1946, the c o n c l u s i o n o f the t r i a l . R e s u l t s and Conclusions:- Owing t o unfavourable weather eon-d i t i o n s , quadrat c l i p p i n g s could not be c a r r i e d out. However, v i s u a l observations confirmed the e a r l i e r r e s u l t s t h a t the value of the p r o t e i n a t e i s i n c r e a s e d markedly by the a d d i t i o n of P 2 0 5 a n d KgO supplements. LAWN TEST: Treatments: N *2°5 KgO 1. M i l l o r g a n i t e 675#/A 40# - -2. (NH 4)gS0 4 200#/A 40# - -3. P r o t e i n a t e pH8.5 500#/A 40# -4. « »t and P 2 0 5 & KgO 650#/A 40# 50# 25# 5. N.P.K. (8-10-5) mix 500#/A 40# 50# 25# 6. C o n t r o l . L o c a t i o n of p l o t and s o i l type: U.B.C. Golf Course, Vancouver, B.C. - Upland G l a c i a l s o i l . T e s t i n g Crop: Mixed grasses, Canada Bl u e g r a s s , Kentucky Bluegrass, Chewing Fescue and Creeping Bent. Design:- Comparative s t r i p s . S i z e of p l o t s : - 10* x 42.5 f Method o f A p p l i c a t i o n : - Surface broadcast. Prooedure:- The t r i a l was l a i d out on A p r i l 26th, 1946 on an area not used e x c e s s i v e l y by the g o l f e r s . Observations were made from time to time and r e s u l t s were based on c o l o r d i f f -erences and v i g o r of growth o n l y . R e s u l t s and Conclusions:- Ammonium Sulphate and N.P.K. mix treatments were n o t i c a b l y b e n e f i c i a l , i n m a i n t a i n i n g d e s i r a b l e c o l o r and vigorous growth. As f o r the remainder of the t r e a t -ments the b e n e f i t was n e g l i g i b l e . - 30 -A STUDY OF THE EFFECT OF PROTEINATE OF SEVERAL pHs, ON THE GERMINATION, SEEDLING DEVELOPMENT,• AND ROOT FORMATION OF SEVERAL PLANT SPECIES Treatments 1. P r o t e i n a t e PH 12 mixed w i t h seed 2. tt PH 10 tt tt tt 3 , n pH 8.5 tt n tt 4 . « • PH 6.0 n « tt . 5-8. tt PH as above §" below seed. 9-12. tt PH tt n 4" beside seed. 13-16. tt PH « tt ^" above seed. 17-20. tt pH tt n side a p p l i c a t i o n 10 days a f t e r germination. 21. C o n t r o l . S o i l : Upland G l a e i a l , • T e s t i n g Seeds: F l a x , Buckwheat, Red Cl o v e r & P e r e n n i a l Ryegrass Design: Randomized block, 2 r e p l i c a t i o n s . Pot S i z e : 5 i " x 3" - (1 l b . s o i l per p o t ) . Procedure: The seeds were placed i n the pots each crop i n a re s p e c t i v e row, t h a t i s four rows o f seeds per pot. The treatments were a p p l i e d as above. The pots were set up outdoors under normal l i g h t and temp-erat u r e c o n d i t i o n s and observations were made d a i l y . R e s u l t s & Conclusions: I t was qu i t e evident t h a t the p r o t e i n -ate ^ r x e g a r d l e s s of the PH and irregajt^LXess of i t s manneroTld^cement, had no d e l e t e r i o u s e f f e c t on the seed germination, development, o r the roo t development. - 31 -A STUDY OF THE ABSORPTION AND MOVEMENT OF PROTEINATE IN  THE SOIL AND ITS RESULTANT EFFECT ON THE VEGETATION SURFACE APPLICATION TESTS  Treatments: ' 1. ^Unjeutr^JLizecl.proteinate pH 12 ) 1 t o 2 ) i n d i l u t i o n s l t o 4 2. N e u t r a l i z e d " pH 8.5) 1 t o 8 S o i l : 1. Outdoor pots - Upland G l a c i a l . 2. E s t a b l i s h e d F i e l d - Sod on Upland G l a c i a l . Method of a p p l i c a t i o n : . Surface spray. Procedure: The treatments were a p p l i e d to the pots and sod on June 1 s t , 1946. The equivalent of 50" of r a i n f a l l was ap p l i e d from June 1 s t to September 1 s t i n order t o a p p r o x i -mate normal l e a c h i n g c o n d i t i o n s . At such a time f i n a l observations were made and conclusions were drawn. R e s u l t s and Conclusions: The p r o t e i n a t e pH 8.5 i n a l l d i l u t i o n s was absorbed completely on both the s o i l and the sod. The p r o t e i n a t e pH 12 i n a l l d i l u t i o n s was not absorbed on the s o i l o r the sod, but appeared to form a c o l l o i d a l membrane (scum) on the su r f a c e . T h i s membrane was i n s o l u b l e . This experimental evidence i n d i c a t e s that the p r o t e i n a t e pH 12 ( l i q u i d ) cannot be absorbed s u c c e s s f u l l y through the surf a c e o f s o i l o r sod and t h e r e f o r e cannot be ap p l i e d as a f e r t i l i z e r through i r r i g a t i o n o r s p r i n k l e r systems. - 32 -A REPORT ON PROTEINATE FERTILIZER IN RELATION TO  BACTERIAL GROWTH This i n v e s t i g a t i o n was undertaken t o determine whether or not s p e c i f i e d p r o t e i n a t e f e r t i l i z e r s were t o x i c to s o i l b a c t e r i a , and to study the u t i l i z a t i o n of the f e r t -i l i z e r n i t r o g e n by these microorganisms.. F e r t i l i z e r s t e s t e d 1. P r o t e i n a t e pH 6.0 - a i r d r i e d meal. 2. P r o t e i n a t e pH 8.5 - a i r d r i e d meal. 3. P r o t e i n a t e pH 10 - a i r d r i e d meal. 4. P r o t e i n a t e pH 12 - a i r d r i e d meal. Composition of F e r t i l i z e r s 20% diatomaceous e a r t h . 8% n i t r o g e n . Rate o f a p p l i c a t i o n o f f e r t i l i z e r s t o s o i l (%), 40 l b . n i t r o g e n per acre. % f e r t i l i z e r i s th e r e f o r e .025% EXPERIMENTS A. To determine the e f f e c t of the p r o t e i n a t e f e r t i l i z e r s on b a c t e r i a l growth and m u l t i p l i c a t i o n . 1. Growth response of pure c u l t u r e s i n the presence of p r o t e i n a t e f e r t i l i z e r s . METHOD An organic medium supporting good growth o f var i o u s s o i l organisms was used. The growth of these organisms i n the b a s a l medium c o n t a i n i n g v a r y i n g amounts of each f e r t i l i z e r I - 33 -was measured and compared t o growth i n the medium without the added f e r t i l i z e r . Growth was measured i n a F i s c h e r Electrophotometer. B a s a l Medium: Ma n n i t o l 1.0% MgS0 4.7H 20 .025% Peptone 2.0% KC1 K 2HP0 4 .025% FeS0 4.7H 20 t r a c e KH 2P0 4 .025% pH 7.2 Organisms t e s t e d : Pseudomonas aeruginosa 9027 R h i z o b i a m e l i l o t i 4602 ' N i t r i f i e r 46-5 N i t r i f i e r 42-5 B a c i l l u s s u b t i l u s 1 F e r t i l i z e r s t e s t e d : #1, 2, 3, and 4 (Page 1)—-' F e r t i l i z e r amounts: 0, .05%, .5% - Water suspensions of each f e r t i l i z e r were made, the pH of each adjusted to 7.2, and the p r e s -c r i b e d amounts o f the f e r t i l i z e r s were added t o the s t e r i l e media a s e p t i e a l l y . P e r i o d of i n c u b a t i o n ; 48 hours, 96 hours. RESULTS Growth measured i n 48 hours. (See next page) - 34 -46-5 42-5 Ps. Aeruginosa Rh. m e l i l o t i N i t r i f i e r N i t r i f i e r B.Subtilus % P e r t : .05% .5% .05% .5% .05% .5% .05%. 5% .05% .5% 15.0 32.5 28.0 27.8 19.5 16.0 15.0 17.5 33.0 29.0 34.0 16.0 21.0 23.0 18.0 35.0 26.0 32.0 16.0 18.0 18.0 18.5 36.0 25.0 22.0 17.0 19.0 16.0 F e r t . f l 44. 41.0 19.0 F e r t . #2 42.0 46.0 20.0 F e r t . fZ 41.0 41.0 21.0 F e r t . jf4 39.0 41.0 19.5 No F e r t i l i z e r 38.0 23.0 33.5 22.5 19.5 Growth measured i n 96 hours: (see Graph 1 also) 46-5 42-5 Ps . Aeruginosa Rh. m e l i l o t i N i t r i f i e r N i t r i f i e r B .Subtilus % F e r t : .05% .5% .05% .5% .05% .5% .05% .5% .05% .5% F e r t . #1 70.0 59.0 23.0 20.0 70.0 31.5 40.0 28.5 22.0 20.5 F e r t . #2 62.0 58.0 22.0 24.5 71.0 29.0 40.0 32.5 23.5 20.5 F e r t . #5 76.5 57.0 22.0 26.0 66.5 28.0 35.0 28.5 22.5 26.0 F e r t . #4 65.0 59.0 23.5 17.0 71.0 25.0 42.5 19.0 22.5 16.0 No F e r t i l i z e r 68.5 23.5 75.0 38.5 22.5 CONCLUSIONS: The f o u r f e r t i l i z e r s gave s i m i l a r r e a c t i o n s at the same l e v e l s . There was no i n h i b i t i o n of any of the organisms w i t h .05% f e r t i l i z e r , or of Ps. aeruginosa w i t h .5% (20 times the normal a p p l i c a t i o n of f e r t i l i z e r ) . However, the higher concentration showed d e f i n i t e i n h i b i t i o n of the n i t r i f i e r s e s p e c i a l l y at 96 hours. Growth of Rh. m e l i l o t i and B. s u b t i l u s was not s u f f i c i e n t to base conclusions upon. - 35 -A s i m i l a r experiment was set up t o determine the amount of f e r t i l i z e r which could be used without decreasing b a c t e r i a l m u l t i p l i c a t i o n . The experiment was a l s o t o show i f continued i n c u b a t i o n would overcome the i n h i b i t o r y e f f e c t of higher concentrations of f e r t i l i z e r . METHOD; Same as previous experiment• B a s a l medium; Same as previous experiment. Organisms; R h i z o b i a leguminosarum - 41-1 N i t r i f i e r 46-5 N i t r i f i e r 42-5 i B a c i l l u s s u b t i l u s F e r t i l i z e r s t e s t e d ; #1, 2, 3, and 4 (Page 1) F e r t i l i z e r amounts; 9, .025$, .05%, 0.1%, 0.2%, 0.3% Water suspensions o f each f e r t i l i z e r were made, the pH adjusted t o 7.2, the suspensions autoclaved, and t h e . pr e s c r i b e d amounts of the f e r t i l i z e r s were added to the s t e r i l e media a s e p t i c a l l y . Time of Incubation; 3 days, 5 days, and 14 days. RESULTS: Tables 1, 2, and 3. Graphs 2, 3, 4, and 5. CONCLUSIONS; I n general, the lower percentages, .025% and .05%, st i m u l a t e d growth oeyond t h a t i n the c o n t r o l s ( b a s a l medium al o n e ) , and the higher percentages, 0.2 and 0.3, i n h i b i t e d growth. The l a t t e r was e s p e c i a l l y t r u e w i t h the n i t r i f i e r s . I n h i b i t i o n was l e s s marked a f t e r two weeks i n c u b a t i o n , but was s t i l l c o n s i d e r a b l e . - 36 -TABLE IX - Growth measured i n 5 days. $ F e r t i l i z e r : 0.0 .025 .05 Rh. leguminosarum: No F e r t : F e r t . # l Fert.#2 Fert.#3 Fert.#4 21.0 N i t r i f i e r 46-5 N i t r i f i e r 42-5 B. S u b t i l u s No.Fert: F e r t . # l Fert.#2 Fert.#3 Fert.#4 No F e r t : Fert.#1 Fert.#2 F e r t ,#Z Fert.#4 No.Fert: F e r t . # l Fert.#2 Fert.#3 Fert.#4 53.5 29.5 22.0 21.5 21.5 26.0 20.0 43.0 41.0 51.0 55.0 25.5 32.5 37.0 31.0 19.5 22.5 20.5 19.0 22.0 25.0 28.0 21.5 41.5 50.0 53.0 53.0 33.0 32.5 33.5 30.0 29.5. 29.0 18.0 20.5 .10 .20 18.5 25.5 18.0 20.0 24.0 20.5 20.0 18.0 43.5 37.5 43.0 39.5 32.0 31.5 32.5 31.5 27.0 29.5 21.0 20.5 TABLE X - Growth measured i n 5 days. % F e r t i l i z e r : 0.0 .025 .05 .10 Rh. leguminosarum: 35.5 32.0 32.0 34.0 32.0 33.0 27.5 26.5 22.5 28.0 20.0 16.0 .20 N i t r i f i e r 46-5 N i t r i f i e r 42-5 B. S u b t i l u s t30 27.0 19.0 20.5 17.0 37.0 29.0 27.0 33.5 30.0 30.0 26.5-27.5 31.0 27.0 21.5 19.5 .30 No. F e r t : 28.5 Fert.#1 28.5 34.0 36.0 32.5 35.0 F e r t .#2 32.5 33.5 30.5 29.5 .25.5 Fert.#3 27.5 27.5 23.0 20.0 20.0 Fert.#4 27.5 32.0. 27.0 21,. 0 24.0 No F e r t : 70.0 F e r t . # l 70.5 66.0 65.0 53.5 46.5 Fert.#2 70.0 68.0 63.0 53.0 46.5 Fert.#3 68.0 58.0 49.5 35.0 . 37.0 Fert.#4 71.0 63.0 55.0 44.0 38.0 No F e r t : 43.0 F e r t . # l 40.5 49.0 43.0 30.0 40.0 Fert.#2 44.5 47.0 38.5 35.0 43.0 Fert.#3 46.0 36.5 34.0 33.0 32.0 Fert.#4 41.5 43.0 41.5 41.0 39.5 No F e r t : 41.0 • F e r t . # l 37.0 38.5 40.0 36.5 41.0 Fert.#2 39.0 41.0 38.5 40.0 33.0 Fert.#3 36.5 35.0 33.5 35.0 40.0 Fert.#4 34.0 37.0 34.5 33.0 30.0 - 37 -TABLE X I - Growth measured i n 14 days % F e r t i l i z e r : 0.0 *025 .05 Rh. leguminosarum: N i t r i f i e r 46-5 N i t r i f i e r 42-5 B. S u b t i l u s No F e r t : F e r t . # l Fert.#2 Fert.#3 Fert.#4 No F e r t : F e r t * # l Fert.#2 Fert.#3 Fert.#4 No F e r t : Fert.#1 Fert*#2 Fert.#3 Fert.#4 No F e r t : F e r t * # l Fert.#2 F e r t ..#3 Fert.#4 36*0 70*0 56.0 43.5 34.0 36.5 32.0 35*0 71*0 74*0 73*0 74*0 75.0 79*0 74*0 60*0 41*0 45.0 43*0 40.0 38.0 32.0 42.0 44.0 68.0 70*0 65.0 76.0 80*0 71.0 75*0 74*0 43.0 40.0 42.5 40*5 .10 .20 34.0 60.0 30.0 18.0 66.0 60.0 61.0 60.0 83*0 79.0 74.0 61*0 49.0 42.0 28.0 36.5 35.0 53.0 24.5 27.5 62.0 65*0 64*0 61*0 79*0 74*0 54.0 65*0 49.0 46.5 38.0 38.5 .30 36.0 45.5 25.0 24.5 53.5 59*0 53.0 60*0 61*0 69*0 64*0 66*0 43.0 47.0 40.0 37.0 2. Growth response of microorganisms i n the s o i l i n the presence of p r o t e i n a t e f e r t i l i z e r s . METHOD: The s o i l s to be t e s t e d were placed i n f l a s k s (200-gnu per f l a s k ) t o which were added d i f f e r e n t percentages of each f e r t i l i z e r . P l a t e counts were c a r r i e d out on the s o i l s , and the growth obtained w i t h the d i f f e r e n t l e v e l s of f e r t i l i z e r was compared to t h a t w i t h c o n t r o l s o i l s c o n t a i n i n g no f e r t i l -i z e r . S o i l s : A. Sandy - pH 6.9 B. Garden- pH 6.5 . . . The pH of the s o i l s was taken before and a f t e r adding the v a r y i n g amounts of each f e r t i l i z e r * No pH changes were detected. * • F e r t i l i z e r s t e s t e d : #1, 2, and 3 (Page 1) F e r t i l i z e r amounts: 0, .025%, and .05% The f e r t i l i z e r meals were added d i r e c t l y t o the s o i l samples without pH adjustments. - 38 -Incubation p e r i o d ; 7 days at room temperature. Medium f o r p l a t e counts: Glucose 1.0 gm. K HPO .5 gm. S o i l e x t r a c t 100 cc Tap water 900 cc RESULTS; T o t a l counts on s o i l s p l u s f e r t i l i z e r s a f t e r 7 days i n c u b a t i o n : (See graph 6 also) SOIL A SOIL B No F e r t i l i z e r 6,400,000 8,600,000 F e r t . # l .025% 14,400,000 9,600,000 .05% 15,000,000 15,900,000 Fert.#2 .025% 11,500,000 . 11,500,000 .05% .21,500,000 29,500,000 Fert.#3 .025% 15,500,000 12,600,000 .05% 24,500,000 19,000,000 CONCLUSIONS: When the s o i l s were p l a t e d f o r t o t a l count a f t e r seven days i n c u b a t i o n , i t was noted that the counts increased p r o p o r t i o n a l l y w i t h the increase i n f e r t i l i z e r . The p r o t e i n -ate m a t e r i a l , by s u p p l y i n g a d d i t i o n a l n i t r o g e n to the s o i l , succeeded i n i n c r e a s i n g the s o i l f l o r a . B. The u t i l i z a t i o n o f f e r t i l i z e r n i t r o g e n by bacteria.". 1. The a b i l i t y of pure c u l t u r e s o f s o i l b a c t e r i a to u t i l i z e the p r o t e i n a t e n i t r o g e n both w i t h and without an a d d i t i o n a l carbon source. METHOD: The microorganisms were grown i n a medium o f s a l t s and mannitol p l u s f e r t i l i z e r , and i n a medium o f s a l t s o n l y p l u s f e r t i l i z e r . A p a r a l l e l experiment i n which the f e r t i l -i z e r was replaced by an equivalent amount of peptone ( i n respect t o nitrogen) was a l s o set up. I n each case, the growth was measured and the production o f ammonia, n i t r a t e , and n i t r i t e , t e s t e d . Medium: M a n n i t o l 1.0% K 2HP0 4 .025% KHgP0 4 .025% MgS0 4.7H 20 .01% KC1 .01% FeS0 4.7H 20 t r a c e Organisms: Ps. Aeruginosa 9027 Rh. leguminosarum 41-1 N i t r i f i e r 46-5 F e r t i l i z e r t e s t e d : #£ pH 8.5 F e r t i l i z e r amounts: F e r t i l i z e r i s 8% N. Added by weight: .025%, .05%, .10% Peptone amounts: Peptone i s 16% N. Added by weight: .0125%, .025%, .05% Tests f o r n i t r o g e n : Ammonia - N e s s l e r ' s reagent N i t r a t e - Diphenylamine N i t r i t e - Tromsdorf's reagent. - 40 -RESULTS: 1. Growth measured i n 4 days: % F e r t i l i z e r : (Ps. aeruginosa With mannitol (Rh. leguminosarum ( N i t r i f i e r tt=5 No mannitol With mannitol No mannitol (Ps. aeruginosa (Rh. leguminosarum ( N i t r i f i e r 4S=B % Peptone: (Ps. aeruginosa (Rh. leguminosarum ( N i t r i f i e r 46-5 (Ps. aeruginosa  (Rh. leguminosarum  ( N i t r i f i e r 46-5 ,025% .05% 13.5 13.0 12.7 13.5 11.0 11.5 19.5 13.0 12.2 14.0 11.0 10.7 14.0 13.5 11.5 19.0 12.0 13.7 .0125% .025% .10% 14.0 11.5 11.0 14.5 11.0 11.0 20.0 13.5 14.0 14.0 12.2 12.2 14.2 12.5 11.7 16.5 14.2 15.2 2. Tests f o r n i t r a t e , n i t r i t e , and ammonia were made i n 48 hours, 3 days, 5 days, and 12 days. However, no n i t r a t e or n i t r i t e were detected at any time; ammonia was found present as f o l l o w s : 48 hours: 3 days: No ammonia M a n n i t o l No mannitol F e r t i l i z e r : .025% P s . aeruginosa 0 Rh. leguminosarum 0 . N i t r i f i e r 46-5 0 Peptone: .012% P s . aeruginosa 0 Rh. leguminosarum 0 N i t r i f i e r 46-5 0 .05% .1% .025% .05% .10% t r a c e t r a c e Trace t r a c e t r a c e 0 0 0 t r a c e t r a c e 0 0 0 t r a c e t r a c e .025% .05% .012% .025% .05% t r a c e t r a c e t r a c e t r a c e t r a c e t r a c e t r a c e 0 p l u s plus 0 0 0 p l u s p l u s - 41 -5 days: M a n n i t o l No M a n n i t o l F e r t i l i z e r : . 025% .05% .10% .025% .05% .10% P s . aeruginosa Rh. Leguminosarum N i t r i f i e r 46-5 0 0-0 0 0 0 t r a c e 0 0 0 0 0 0 p l u s 0 p l u s p l u s p l u s Peptone: .012% .025% .05% .012% .025% .05% Ps. aeruginosa Rh. leguminosarum N i t r i f i e r 46-5 0 0 0 0 0 0 t r a c e 0 0 0 p l u s •o 0 p l u s .. t r a c e t r a c e p l u s t r a c e 12 days: F e r t i l i z e r : P s . aeruginosa  Rh. leguminosarum N i t r i f i e r 46-5 M a n n i t o l No M a n n i t o l ,025% .05% .10% .025% .05% 0 0 t r a c e p l u s p l u s 2 p l u s 2 0 0 0 p l u s p l u s 2 p l u s 2 0 0 0- p l u s p l u s 2 p l u s 2 Peptone: Ps. aeruginosa Rh. leguminosarum N i t r i f i e r 46-5 .012% .025% .05% 0 0 t r a c e 0 0 t r a c e 0 0 0 ,012% .025% .05% p l u s p l u s 2 p l u s p l u s 2 p l u s pius 2 p l u s 2 p l u s 2 p l u s 2 CONCLUSIONS: The organisms used were capable of u t i l i z i n g the pro-t e i n a t e f e r t i l i z e r f o r carbon and n i t r o g e n e q u a l l y as w e l l as they could the peptone. Meagre growth was obtained i n a l l cases because i n the presence of mannitol, n i t r o g e n was the l i m i t i n g f a c t o r , and i n the absence of m a n n i t o l , carbon was the l i m i t i n g f a c t o r . Ammonia was relea s e d from both the peptone and the f e r t i l i z e r . I n the presence of a d d i t i o n a l carbon and energy - 42 -source ( m a n n i t o l ) , the ammonia was u t i l i z e d by the organisms. However, i n the absence o f mannitol, the ammonia accumulated. Therefore, the organisms t e s t e d were found able t o u t i l i z e the p r o t e i n a t e n i t r o g e n e q u a l l y as w e l l as t h a t of pep-tone. 2. The a b i l i t y of the s o i l f l o r a to u t i l i z e the p r o t e i n a t e n i t r o g e n . METHOD and RESULTS: 200 gm. samples o f s o i l s c o n t a i n i n g .025% and .05% f e r t i l i z e r were allowed t o incubate a t room temperature f o r two weeks. At the end of t h i s time, no t r a c e of ammonia, n i t r a t e , or n i t r i t e could be detected. CONCLUSIONS: The carbon n i t r o g e n r a t i o i n the s o i l was such t h a t the organisms were s u p p l i e d w i t h s u f f i c i e n t energy t o u t i l i z e t h e ni t r o g e n p r e s e n t . SUMMARY AND CONCLUSIONS The p r o t e i n a t e f e r t i l i z e r s , when added t o the s o i l a t the r a t e o f 40 l b s . of n i t r o g e n per ac r e , act as a source of n u t r i e n t s to the b a c t e r i a and hence tend t o increase the s o i l f l o r a . I n the presence of a s u i t a b l e carbon source, micro-organisms r e l e a s e ammonia from the f e r t i l i z e r s and u t i l i z e the n i t r o g e n f o r c e l l s y n t h e s i s . - 43 -A COMPARISON OF THE EFFECT OF LEACHING ON PROTEINATE WITH THE EFFECT ON COMMERCIAL FERTILIZERS LABORATORY TRIAL  S o i l s : 1. I n t e r i o r 2. Upland G l a c i a l 3. Lowland Treatments: Rate of A p p l i c a t i o n 1. C o n t r o l 2. P r o t e i n a t e pH 12 ( l i q u i d ) 5 gms. 3. 4-10-5 5 gms. 4. P r o t e i n a t e pH 6 ( s o l i d ) R e p l i c a t i o n s : two Prooedure: 2.5# s o i l were placed i n g l a s s tubes 18" x 1.5" The treatments were incorporated i n the s o i l . Water was allowed t o drop i n t o the tubes a t the rat e of 5.8 cc. per hour, per day - t o t a l l i n g 1690.4 c c . T h i s was e q u i v a l e n t to 100" of p r e c i p i t a t i o n . The l e a c h i n g s were caught under the tubes i n f l a s k s and were kept under constant evaporat-i o n i n order to m a i n t a i n a workable volume. The concentrated l e a c h i n g s were then analyzed f o r n i t r a t e n i t r o g e n and con-c l u s i o n s drawn from the r e s u l t s . Conclusions: The r e s u l t s were not very c o n c l u s i v e and few v a l i d statements could be made. However, i t may g e n e r a l l y be s a i d t h a t p r o t e i n a t e pH 12 and pH 6 l e a c h through the s o i l as r e a d i l y as commercial f e r t i l i z e r s . - 44 -A COMPARISON OF THE EFFECT OF LEACHING ON  PROTEINATE ALL pH CONCENTRATIONS FIELD TRIAL  S o i l : Upland G l a c i a l Treatments: -1. P r o t e i n a t e pH 12 Incorporated 3.4 gms. (6C#N/A) 2. " " Surface 3. " pH 10 Incorporated 4. " " Surface 5. " pH 8.5 Incorporated 6. » • " Surface 7. " 1 Ph 6.0 Incorporated 8. n n Surface 9. C o n t r o l N I L R e p l i c a t i o n s : Each treatment was d u p l i c a t e d . 18 treatments were used outdoors and 18 i n d o o r s . Procedure: 20# of s o i l were placed i n each of the 36 pots and the r e s p e c t i v e treatments were a p p l i e d . A d u p l i c a t e d set of pots was p l a c e d outdoors and thus exposed t o the weather. The other d u p l i c a t e d set of pots was placed indoors thus r e c e i v i n g no e f f e c t s from the weather. The outdoor pots were set out on February 21st, 1946 and l e f t u n t i l September 21st, when a l l pots outdoors and indoors were set up on the green-house bench and seeded w i t h Storm Rye. The t e s t crop was harvested on November 24th and Green top weights were r e c o r -ded. The tops were then oven d r i e d and the. d r y weights recorded. From"these r e s u l t s , conclusions were drawn. - 45 -TABLE X I I - F i e l d Leaching T r i a l R e s u l t s GREENHOUSE DESIGN pH 6 Inc. pH 10 S u r f . pH 12 S u r f . (D pH 6 Inc. (L) pH6 S u r f . PHIO : [nc. pH 10 Inc. (L) pH 6 S u r f . (L) pH 8.5 S u r f . pH 10 Surf. (L) pH 12Inc. (L) C o n t r o l (L) pH 6 S u r f . (L) pHIO S u r f . pH 8.5 I n c . PH8.5 Su r f . pH8.5 Inc PH12 Surf " (D pH 12 I n c . (L) pH12 I n c . C o n t r o l pH 12 S u r f . pHIO Su r f (L) i pH6 ] :nc. pH 6 S u r f . pHIO I n c . pH 8.5 S u r f . (L) pH 8.5 I n c . (L) pH 6 I n c . (L) pHIO (L) Inc C o n t r o l (D pH12 I n c . C o n t r o l pH 12 S u r f . pH8.5 Inc (L) S u r f pH8.5 (D DRY WEIGHT RESULTS (Green Wts.) 10.2 gm. 82.5 7.4 gm. 62.7 2.5 gm. 14.6 2.6 gm. 18.3 9.4 gm. 61.6 8.8 gm. 52.3 3.2 gm. 17.0 1.8 gm. 16.0 7.1 gm. 60.8 1.9 gm. 11.4 2.7 gm. 23.05 2.7 gm. 19.2 2.5 gm. 18.8 9.2 gm. 68.2 6.5 gm. 41.5 9.1 gm. 62.7 7.4 gm. 45.6 2.6 gm. 15.05 2.4 gm. 21.8 7.8 gm. 7.6 gm. 30.6 7.5 gm. 72.5 2.1 gm. 13.6 9.8 gm. 76.4 10.1 gm. 70.8 9.8 gm. 60.2 2.6 gm. 14.4 3.2 gm. 19.4 2.8 gm. 20.1 2.8 gm. 14.2 1.8 gm. 18.4 8.2 gm. 78.3 6.6 gm. 29.4 7.8 gm. 74.2 2 .8 gm. 16.8 2.2 gm. 14.3 CONCLUSIONS From both t he leached r e s u l t s and the unleached r e s u l t s , i t was not4d t h a t the inco r p o r a t e d treatment gave g r e a t e r y i e l d s than the surface treatment, w i t h the exception of pH 8.5 i n the unleached p o t s . Thus i t might be s a i d t h a t surface a p p l i c a t i o n s of the p r o t e i n a t e would not be a d v i s a b l e as the r a t e o f l e a c h i n g i s too gr e a t . As a ge n e r a l c o n c l u s i o n p r o t e i n a t e at a l l pHs le a c h t o a considerable extent but as proven i n the previous experiment to no gre a t e r . e x t e n t than the commercial f e r t i l i z e r s . - 47 -A DETERMINATION OP THE MINOR ELEMENT CONTENT OP PROTEINATE 20 gms. of the p r o t e i n a t e powder pH 8 . 5 was ashed i n an i n c i n e r a t o r and a speetographic a n a l y s i s was c a r r i e d out on t h i s ash. This experiment was performed t o determine what minor elements were present i n the m a t e r i a l and the app-roximate r a t i o s of the var i o u s elements. The r e s u l t s obtained must not be taken as representing a q u a l i t a t i v e a n a l y s i s . RESULTS / Elements not Band Reading I n d i c a t e d Present 20 P r i n c i p a l C o n s t i t u e n t s Sodium Magnesium S i l i c a Other Elements Present 10 10 Phosphorous 6 Calcium 4 Aluminium 2 Copper 2 L i t h i u m 2 Potassium 1 Lead 1 T i n 1 S i l v e r .01 I r o n .2 Zi n c .2 Mercury .001 Barium .001 Cobalt Molybdenum. Boron Cadmium Sulphur Manganese A r s e n i c P a l l a d i u m N i c k e l Titamium - 48 -Bismuth Strontium CONCLUSION: I n s o f a r as minor elements are u s u a l l y added to f e r t i l i z e r s as r e q u i r e d , i t would appear th a t -the p r o t e i n a t e s a t i s f i e s the minor element requirements w i t h the exception of Boron, Manganese, and Sulphur which a r e not i n d i c a t e d as being p r e s e n t . On the b a s i s o f t h i s spectographic a n a l y s i s , i t i s i n d i c a t e d t h a t a f u r t h e r determination o f minor elements by a q u a n t i t a t i v e method would be worthy o f c o n s i d e r a t i o n . .001 .1 AN INVESTIGATION AS TO THE SUITABILITY OF PROTEINATE  TO PASS THROUGH THE STANDARD FERTILIZER APPLICATIONS Method On February 14th, 1946 the p r o t e i n a t e pH 8.5 (dry powder w i t h a diatomaceous e a r t h content of 20%) was t e s t e d f o r s u i t a b i l i t y of a p p l i c a t i o n t o the l a n d through standard d r i l l s and a p p l i c a t o r s . . I t was found t h a t w h i l e the m a t e r i a l passes through the adaptors s a t i s f a c t o r i l y owing t o the small p a r t i c l e s i z e , i t would not run through the d e l i v e r y tubes i f the angle i s g r e a t e r than 10 degrees from the v e r t i c l e owing to the l i g h t and deliquescent nature of the m a t e r i a l . Another problem that arose from t h i s t r i a l was the ease w i t h which the p r o t e i n a t e d r i f t e d when the wind was blowing even t o the s l i g h t e s t e xtent. CONCLUSIONS This m a t e r i a l must be converted t o a g r a n u l a r form. I n t h i s way t h e r e w i l l not be the l o s s of f e r t i l i z i n g value through d r i f t i n g and g r a n u l a t i o n w i l l a s s i s t i n a l l o w i n g the m a t e r i a l to pass through the a p p l i c a t o r s a t g r e a t e r angles. A l s o , the problem of deliquescence must be overcome f o r ease i n a p p l i c a t i o n and ease of h a n d l i n g and s t o r i n g . - 50 -INVESTIGATIONS INTO THE PROBLEM 0? PROTEINATE DELIQUESCENCE  Container Test: M a t e r i a l s : -1. P r o t e i n a t e pH 10 i n diatomaceous e a r t h . 2. P r o t e i n a t e pH 8.5 » » " . 3. P r o t e i n a t e pH 6.0 " " " . Containers:-1. C l o t h Bags. 2. T r i p l e K r a f t Paper Bags. 3. K r a f t Laminated Asphalt Bags. Procedure: The moisture content of the v a r i o u s p r o t e i n a t e samples were determined and the f o l l o w i n g r e s u l t s were obtained: P r o t e i n a t e pH 10 2.58% P r o t e i n a t e 8.5 1.79% P r o t e i n a t e 6.0 2.00% The samples were then placed i n the d i f f e r e n t types of con-t a i n e r s as noted above, and l e f t under dry storage c o n d i t i o n s . Observations were made from time t o time and w i t h i n a month's time the samples i n the C l o t h and T r i p l e K r a f t containers had to be discarded i n t h a t the m a t e r i a l had taken on so much water that i t was running out through the already r o t t e d c o n t a i n e r s . As f o r the K r a f t Laminated Asp h a l t c o n t a i n e r , the samples were l e f t i n the bags u n t i l the f o l l o w i n g year when they were removed and a moisture content run on the m a t e r i a l s . The r e s u l t s of t h i s a n a l y s i s were as' f o l l o w s : - 51 -P r o t e i n a t e pH 10 P r o t e i n a t e pH 8.5 P r o t e i n a t e pH 6.0 CONCLUSIONS I n t h a t the average moisture. content of the o r i g i n a l samples was 1.79% and a f t e r one year i n storage w i t h the Laminated Asphalt c o n t a i n e r the average moisture content was 5.28%-, only 3.49% moisture had been absorbed. Thus, p r o t e i n -ate cannot be stored i n the standard f e r t i l i z e r c o n t a i n e r s but must be stored i n a Laminated As p h a l t Bag or some other c o n t a i n e r which w i l l prevent an excess o f a i r c o n t a c t i n g t he m a t e r i a l . The experiment a l s o i n d i c a t e s , once again, t h a t the m a t e r i a l i s very much too d e l i q u e s c e n t . Some method o f meeting t h i s d i f f i c u l t y must be devised before e f f e c t i v e use can be made o f p r o t e i n a t e as a f e r t i l i z e r . 5.63% 4.87% 5.36% - 52 -A METHOD .OF PREPARING A COMPLETE N.P.K.  FERTILIZER WITH THE PROTEINATE BASE -P r e v i o u s l y performed experiments i n d i c a t e d t h a t when supplemented w i t h Pg^s K2°» t J l e p r o t e i n a t e exerted a more b e n e f i c i a l e f f e c t on p l a n t growth. I t has a l s o been concluded th a t a granular form would e r a d i c a t e the d i f f i c u l t y of the m a t e r i a l being too l i g h t and powdery f o r the standard f e r t i l -i z e r a p p l i c a t o r s . With these p o i n t s i n mind, the f o l l o w i n g method a f t e r c o n s i d e r a b l e experimentation, was found t o be a move i n the r i g h t d i r e c t i o n . M a t e r i a l s : 1. P r o t e i n a t e pH 12 8# a t 8% N 2. Superphosphate 5# a t 18% P 2 0 5 3. Muriate of potash 1# at 60% K 20 Procedure: The p r o t e i n a t e was placed i n a mixing bowl and the superphosphate was added w i t h mixing u n t i l the mass was o f uniform c o n s i s t e n c y . The muriate of potash was then added and the mixture once again s t i r r e d completely. The r e s u l t a n t sludge was placed i n a c l o s e d c o n t a i n e r and allowed to cure f o r a p e r i o d o f two months under normal temperature c o n d i t i o n s . At t h i s time the m a t e r i a l was exposed t o the a i r and consequen-t l y d r i e d w i t h i n a matter' o f days. The d r i e d m a t e r i a l was then placed i n a g r i n d e r and ground to 200 mesh p a r t i c l e s i z e ; I n t h i s regard i t might be w e l l to note t h a t t h i s p a r t i c l e s i z e could e a s i l y be a l t e r e d i f i t be necessary. R e s u l t s : The product of the aforementioned procedure was found to he n o t a b l y l a c k i n g i n i t s d e l i q u e s c e n t p r o p e r t i e s as i t maintained i t s d r y g r a n u l a r form on exposure to the a i r . This form of m a t e r i a l from mechanical standpoints i s as good, i f not b e t t e r than the standard organic mixes of approximately the same f o r m u l a t i o n i n t h a t i t i s not so l i k e l y to d r i f t w i t h the wind and w i l l f l o w e a s i l y through f e r t i l i z e r a p p l i c a t o r s . - 54 -DETERMINATION OF THE PROTEIN INDEX OF THE PROTEINATE To some extent the n u t r i t i v e values of animal p r o t e i n concentrates have been i n v e s t i g a t e d from the standpoint o f type and c o n d i t i o n o f raw m a t e r i a l s , method of cooking, and methods o f d r y i n g and some explanations have been advanced as to the causes o f v a r i a t i o n i n n u t r i t i v e v a l u e s . F o r example, i t has been shown t h a t the e d i b l e p o r t i o n s of f i s h are sup e r i o r to and more a i g e s t a b l e than meal prepared from cann-ery f i s h waste (11). A method o f determining the n u t r i t i v e v a l u e of animal p r o t e i n s by chemical means has been developed (1) . T h i s method i s v e r y r a p i d when compared to b i o l o g i c a l t e s t s which take considerable time. I t i s r e l a t i v e l y simple and i s l i m i t e d to the determination of those forms of n i t r o g e n whieh are considered to have the g r e a t e s t i n f l u e n c e on n u t r i t i v e v a l u e . The p r o t e i n index i s determined by t h i s method and the a n a l y t i c a l d e t a i l s are g i v e n below. The p o t e n t i a l p o s s i b i l i t i e s of p r o t e i n a t e as a feed have been i n v e s t i g a t e d and to a degree have proved r a t h e r promising. Y e t , to the authors' knowledge, the t e s t s thus f a r c a r r i e d out have been l i m i t e d i n scope. As s t a t e d e a r l i e r i n t h i s paper, i t i s not our purpose to make a d e t a i l e d i n v e s t i -g a t i o n of the f e e d i n g value of p r o t e i n a t e , but r a t h e r , to make a few t e s t s as an i n d i c a t i o n of the v a l u a b l e i n f o r m a t i o n which might be obtained i n f u t u r e s t u d i e s . METHODS USED IN THE FRACTIONATION OF THE NITROGEN IN ANIMAL PROTEIN CONCENTRATES Reagents Magnesium oxide Alum s o l u t i o n Copper sulphate s o l u t i o n 100 gm. CUSO4.5H2O per l i t e r 100 gm. N a A l ( S 0 4 ) 2 . 1 2 H 2 0 " 50 gm. MgO suspended i n 1 " H y d r o c h l o r i c a c i d 250 gm. H c l i n 1 l i t e r P e p s i n s o l u t i o n 5 gm. pepsin i n 1 l i t e r Phosphotungstic a c i d Acetate b u f f e r Approximately l m o l a l , pH Approximately 5. Procedure: A Quantity of the concentrate c o n t a i n i n g 0.5 to 1.0 gm. of crude p r o t e i n was placed i n a porous alundum c r u c i b l e and extr a c t e d w i t h e t h y l ether f o r approximately 24 hours i n a Bailey-Walker apparatus. I t was then t r a n s f e r r e d to a 250 c c . beaker, mixed w i t h 80 cc. of water and heated on a steam bath f o r 30 minutes. F i v e cubic centimeters of alum reagent, 25 cc . of copper sulphate s o l u t i o n and 50 c c . of magnesium oxide suspension were then added. Heating on a steam bath was continued f o r 20 minutes l o n g e r . When c o o l , the p r e c i p i t a t e was f i l t e r e d o f f and washed three times w i t h 25 c c . of 2% copper sulphate s o l u t i o n . The n i t r o g e n i n the p r e c i p i t a t e was determined. f o r 30 minutes w i t h a s l i g h t excess of magnesium oxide to The f i l t r a t e and washings from the above were b o i l e d - 56 -d r i v e o f f ammonia and amines, f i l t e r e d , washed and made a c i d w i t h h y d r o c h l o r i c a c i d to about 1 N. Ten cubic centimeters of phosphotungstic a c i d were added and the s o l u t i o n was kept i n an i c e box over n i g h t . More phosphotungstic-. a c i d was added i f p r e c i p i t a t i o n had not been complete, but a l a r g e excess was avoided. The p r e c i p i t a t e was f i l t e r e d o f f , u s i n g a f i l t e r a i d i f necessary, washed once w i t h i c e c o l d 1 normal h y d r o c h l o r i c a c i d c o n t a i n i n g about 1% of phosphotungstic a c i d and the n i t r o g e n content of the p r e c i p i t a t e determined. The u n d i g e s t i b l e p r o t e i n was determined on a f r e s h 2 gm. sample which was e x t r a c t e d w i t h e t h y l ether as before and t r a n s f e r r e d to a 500 c c . shaking b o t t l e . Two hundred and f i f t y cubic centimeters of water, 5 c c . of h y d r o c h l o r i c a c i d and 25 cc. of f r e s h l y prepared pep s i n s o l u t i o n were added. The mixture was shaken f o r 25 hours a t a temperature of 37 degrees C. Then 5ce. of h y d r o c h l o r i c a c i d were added and the mixture was allowed t o stand f o r 24 hours more at a temperature of 37 degrees C. The i n s o l u b l e m a t e r i a l was f i l t e r e d and washed.two or three times w i t h hot water. To o b t a i n c l e a r f i l t r a t e s , a f i l t e r a i d was o c c a s i o n a l l y necessary. The n i t r o g e n i n the undissolved residue was determined. The hot water s o l u b l e p r o t e i n was determined by e x t r -a c t i n g a 2 gm. sample w i t h e t h y l ether u n t i l most of the f a t was removed, then b o i l i n g w i t h 200 c c . of water c o n t a i n i n g - 57 -5 cc. of the acetate b u f f e r f o r 30 minutes. The mixture was f i l t e r e d w h i l e hot and washed s e v e r a l times w i t h hot water. The f i l t r a t e and washings were t r e a t e d w i t h c u p r i c i o n and magnesium oxide as al r e a d y described, and the n i t r o g e n content of the copper p r e c i p i t a t e determined. APPARATUS FOR DETERMINATION OF UNDIGESTIBLE PROTEIN ILLUSTRATION 8 R e s u l t s : 1. Copper ppt. contained .105 gm. N i t r o g e n - 50.2% 2. Hot water S o l u b l e .049. gm. » - 23.4% 3. Phosphotungstic .031 gm. " - 14.8% 4. Undigestable .014 gm. " - 6.6% P r o t e i n Index: 50.2 - (6.6 p l u s .6 x 23.4) p l u s .4 X 14.8 r 35.48 Conclusion: From the above r e s u l t s we are able t o s t a t e t h a t p r o t e i n a t e has a p r o t e i n q u a l i t y index of 35.48. I n compar-i s o n w i t h r e s u l t s obtained by H.J. Almquist ( 1 ) , t h i s mater-i a l has a lower index than any of the animal p r o t e i n w i t h which he experimented. However, the r e s u l t s compared q u i t e f a v o u r a b l y w i t h the d o g f i s h meal a n a l y s i s being o n l y a l i t t l e lower i n the v a r i o u s percentages. The a n a l y t i c a l procedure which was employed, w h i l e i t does not detect the presence of t o x i c substances, does permit of the c o n c l u s i o n t h a t there i s a good p o s s i b i l i t y of t h i s m a t e r i a l being used as a p r o t e i n supplement f e e d . This c o n c l u s i o n should be s u b s t a n t i a t e d w i t h adequate b i o l o g i c a l t e s t s . - 59 -A STUDY PIT THE EFFECT OF PROTEINATE ON THE PAL AT ABILITY OF VARIOUS FODDER CROPS Procedure: F o l l o w i n g the p r e v i o u s l y o u t l i n e d experiment on the e f f e c t o f p r o t e i n a t e as a f e r t i l i z e r on pa s t u r e , c a t t l e were turned i n t o the p l o t area and were allowed to feed on the t r e a t e d p l o t s . A check was made on the g r a z i n g h a b i t s of the . c a t t l e to determine whether o r not preference was shown f o r the untreated, areas. Conclusion: • -I t was noted that the c a t t l e found the pasture as p a l a t a b l e where p r o t e i n a t e had been a p p l i e d as where i t had not been a p p l i e d . There was l i t t l e evidence of the t r e a t -ments on the p l o t s a t t h i s time as most of i t had e i t h e r been absorbed by the p l a n t s o r had leached away through the s o i l . I t i s d o u b t f u l t h a t p r o t e i n a t e would cause l i v e s t o c k t o a v o i d t r e a t e d areas t o any g r e a t e r extent than would be caused by any organic f e r t i l i z e r a t a s i m i l a r time a f t e r a p p l i c a t i o n . - 60 -DISCUSSION From the experimental' work c a r r i e d out on t h i s by-product, c e r t a i n advantages and disadvantages o f the m a t e r i a l have been made evide n t . As a f e r t i l i z e r , the p r o t e i n a t e has the advantage i n t h a t i t i s an a v a i l a b l e source of n i t r o g e n which may be relea s e d g r a d u a l l y as a p l a n t food. I t has no harmful e f f e c t on s o i l microorganisms but r a t h e r tends to st i m u l a t e t h e i r a c t i v i t y ; i t has no adverse e f f e c t on seed germination o r r o o t development. The disadvantages o f t h i s m a t e r i a l as a f e r t i l i z e r are many. The extreme deliquescence together w i t h the powdery form of the p r o t e i n a t e r e s u l t i n t h i s m a t e r i a l being extremely d i f f i c u l t t o apply through the standard f e r t i l i z e r machinery. I n a d d i t i o n , i t i s r a t h e r obnoxious t o handle and does not stand up under storage c o n d i t i o n s . I t might be s a i d t h a t the disadvantages of the pr o t e i n a t e outweigh the advantages, but i t has a l s o been noted t h a t these disadvantages may be reduced to a minimum at reasonable cost and the f i n a l product would have a h i g h standard as a commercial f e r t i l i z e r . I t has a l s o been noted t h a t the l i q u i d form of the p r o t e i n a t e has a s o i l p a r t i c l e aggregating e f f e c t which might prove most u s e f u l i n the c o a g u l a t i o n of d e f l o c u l a t e d p r a i r i e s o i l s . The e f f e c t s on s o i l s t r u c t u r e have not been i n v e s t i -gated i n t h i s paper t o any extent,.but i t i s the authors* b e l i e f t h a t f u r t h e r s t u d i e s along t h i s l i n e would be sound and t i m e l y as an a i d to s o i l conservation p r a c t i c e s . - 61 r As a feed supplement, the P r o t e i n Index obtained i n d i c a t e s t h a t t h i s m a t e r i a l may have some use i n animal r a t i o n s . On the b a s i s of the r e s u l t s obtained i n t h i s i n v e s t i g a t i o n , the m a t e r i a l cannot be recommended as a l i v e -stock feed. However, there are i n d i c a t i o n s t h a t should f u r t h e r t e s t s of a b i o l o g i c a l nature be performed, they would be of value and might p o s s i b l y l e a d t o the development o f a new p r o t e i n feed supplement. I n view o f the present day c o n d i t i o n s as to the supply of commercial organic nitrogenous f e r t i l i z e r s , i t may be s t a t e d that the p r o t e i n a t e has a p l a c e i n the f e r t i l i z e r i n d u s t r y . I t would be a d i s t i n c t advantage t o B r i t i s h Columbia should the shortcomings of the p r o t e i n a t e be remedied and the m a t e r i a l placed on a sound economic p r o d u c t i o n b a s i s and made a v a i l a b l e t o the 'organic hungry' f e r t i l i z e r consumers of today. - 62 -CONCLUSIONS FERTILIZER VALUE 1. The p r o t e i n a t e i n powder form shows no s i g n i f i c a n t d i f f e r e n c e between the v a r i o u s pHs as a f e r t i l i z e r . 2. The p r o t e i n a t e i n powder form i s equal to commercial organic f e r t i l i z e r s but i s hot equal to commercial i n o r g a n i c f e r t i l i z e r s . 3. I n l i q u i d form, the p r o t e i n a t e does not compare favour-a b l y w i t h any commercial f e r t i l i z e r . 4. The p r o t e i n a t e , i n a l l forms, leaches from the s o i l but to no g r e a t e r extent than commercial f e r t i l i z e r s . 5. The l i q u i d p r o t e i n a t e cannot be absorbed through the surface of e i t h e r s o i l or sod. 6. The p r o t e i n a t e a c t s as a source of n u t r i e n t s t o s o i l b a c t e r i a and tends t o increase the s o i l f l o r a . I n the presence o f a s u i t a b l e carbon source, microorgan-isms r e l e a s e ammonia from the p r o t e i n a t e and u t i l i z e the n i t r o g e n f o r c e l l s y n t h e s i s . 7. When the p r o t e i n a t e i s incorporated i n the s o i l , g r e a t e r y i e l d s are obtained than when surface a p p l i -c a t i o n s are made. 8. With the exception of Boron, Manganese, and Sulphur, which are not i n d i c a t e d as being present, the p r o t e i n -ate meets minor element requirements. 9. The p r o t e i n a t e i n powder form, owing t o the small p a r t i c l e s i z e and l i g h t d e l i q u e s c e n t nature, cannot be a p p l i e d through standard f e r t i l i z e r machinery and must be converted to some granular form. 10. I t i s p o s s i b l e to make a complete N.P.K. f e r t i l i z e r having a base of the l i q u i d p r o t e i n a t e . T h i s m a t e r i a l i s g r a n u l a r i n form and deliquescence i s markedly reduced. FEEDING VALUE 1. The p r o t e i n a t e index i s lower than t h a t of common animal p r o t e i n feed supplements but shows some value s u f f i c i e n t t o warrant f u r t h e r experimental work. 2. When the p r o t e i n a t e i s a p p l i e d as a f e r t i l i z e r to forage crops, there i s no adverse e f f e c t on the p a l a t a b i l i t y o r n u t r i t i v e value of such crops. - 63 -LITERATURE CITED Almquist, H.J. Stoks t a d , E.L.R. Halbrook, E.R. B a r t l e t t , J.M. Broklesby, H.N, e t . a l . Brown, F.W. Jenk i n s , E.H. S t r e e t , J.P. Le C l e r c , J.A. Ross, Wm.H. Stevenson, C.H. Tu r r e n t i n e , J.W. "Supplementary Values of Animal P r o t e i n Concentrates, i n Chick Rations" J . N u t r i t i o n , Vol.10, No.2, Page 193. " F i s h Wastes For Feeding Animals" Marine A g r . E x p . S t a . B u l l . 266. "The Chemistry and Technology of Marine Animal O i l s With P a r t i c -u l a r Reference t o Those of Canada" F i s h Res.Bd.Can. B u l l . 59. "The Sources of Our Nitrogenous F e r t i l i z e r s " U.S.D.A. B u l l . 729 (1917). "Manure From The Sea" Con. Agr. Exp. S t a * B u l l . 194 "The F i s h Meal Industry" U.S. Bur.For. and Dom.Commerce B u l l . 538. " F e r t i l i z e r s From I n d u s t r i a l Wastes" U.S.D.A. B u l l . 728 (1917) "Aquatic Products As F e r t i l i z e r s " U.S. Comm. of F i s h and F i s h e r i e s 1902, Page 253. " U t i l i z a t i o n of the F i s h Waste Of The P a c i f i c Coast For the Manu-f a c t u r e of F e r t i l i z e r " . U.S.D.A. B u l l . 150 (1915). - 64 -(10) Weber, P . O . (11) W i l d e r , O.H.M. Bethke, P.M. Record, P.R. " F i s h Meal: I t s Use As A Stock And P o u l t r y Food" U.S.D.A. B u l l . 378 (1916). " E f f e c t of Method of Manufacture on the- N u t r i t i v e Value of F i s h Meal as Determined by N i t r o g e n Balance s t u d i e s w i t h Rats" J . Ag. Res. V o l . 49, Page 723. 

Cite

Citation Scheme:

        

Citations by CSL (citeproc-js)

Usage Statistics

Share

Embed

Customize your widget with the following options, then copy and paste the code below into the HTML of your page to embed this item in your website.
                        
                            <div id="ubcOpenCollectionsWidgetDisplay">
                            <script id="ubcOpenCollectionsWidget"
                            src="{[{embed.src}]}"
                            data-item="{[{embed.item}]}"
                            data-collection="{[{embed.collection}]}"
                            data-metadata="{[{embed.showMetadata}]}"
                            data-width="{[{embed.width}]}"
                            async >
                            </script>
                            </div>
                        
                    
IIIF logo Our image viewer uses the IIIF 2.0 standard. To load this item in other compatible viewers, use this url:
http://iiif.library.ubc.ca/presentation/dsp.831.1-0106964/manifest

Comment

Related Items