P H Y S IC O -C H E M IC A L C H A R A C T E R IS T IC S O F SHRIM P FEEDS COMPOUNDED FROM A FEW FER M E N TED FEED IN G R E D IE N TS
DISSERTATION SUBMITTED
IN P A R T IA L FULFILM ENT OF THE REQUIREMENTS FOR THE DEGREE OF
MASTER OF FISHERIES SCIENCE (MARICULTURE)
OF THE
C E N T R A L INSTITUTE OF FISHERIES E D U C A T I O N (DEEMED UNIVERSITY)
BY SADU HARI
» .SC'-
... - ...
CENTRAL MARINE FISHERIES RESEARCH INSTITUTE
(INDIAN COUNCIL OF AGRICULTURAL RESEARCH) C O C H IN -6 8 2 014
INDIA.
JULY 2000
Dedicated to
my beloved parents
C E R T I F I C A T E
Certified that the disserialion entitled " Physico-chemical characteristics of shrimp feeds compounded from a few fermented feed ingredients " is a bonafide record of work done by Mr. Hari, S. under our guidance at the Central Marine Fisheries Research Institute during the tenure o f his M. F. Sc. (Mariculture) programme (1998-2000) and that it has not previously formed the basis for the award of any other degree, diploma or other similar titles or for any publication.
Dr. (Mrs.) Manpal Sridhar (Chairman & Major Adviser),
Scientist (S. S), CMFRI, Cochin,
(Co-Chairman), Head, PNiMX CMFRK Cochin.
'ijayagopal, (Member),
Scientist, CMFRl, Cochin
/
V, n / ^ Dr. M. Srinath.
(Member).
Sr Scientist, CMl Rl, Cochin.
D EC L A R A T IO N
I hereby certify that this thesis entitled "Physico -chemical characteristics o f shrimp feeds com pounded from a few fermented feed ingredients" is based on my own research and has not prev io u sly formed the basis for the aw ard o f any degree, diploma, associateship, fellowship or other similar titles o f recounition.
Place: Cochin Sadu Hari
CONTENTS
Page no.
• List o f Figures, Plates, Tables I-V
•A cknow ledgem ent Vl-VII
•Introduction 1-3
•R e v ie w o f Literature 4-11
•M aterials A nd Methods 12-20
•R esults 21-36
•D iscussion 37-41
•S u m m a ry 42-44
•R eferences 45-49
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Figure I. H y d ro s ta b ility o f feeds co m p o u n d ed with u n ferm ented dried cab b ag e waste (CW ).
Figure 2. H y d ro s ta b ility o f feeds co m pounded with dried cab b ag e w aste ferm ented using B a cillu s c o o g u la n s.
Figure 3. H y d ro s ta b ility o f feeds c o m p o u n d e d with dried cab b ag e w aste ferm ented using B e a u v eria sp.
Plate I; I rcsh c a b b a g c w a s te and cooked, dried and pow dered cabbage waste.
Plate II: Fresh c u ltu re o i B a c illm co a g u la n s used for solid state ferm entation o f cabbagc w aste (C W ), C o tto n s e e d cake (C S C ) and Gingelly oilcake (GOC).
Plate III: D rie d a n d p o w d e r e d cottonseed cake and gingelly oilcake b efore inoculation w ith b a c te r ia a n d fungi for fermentation.
Plate IV: T h e c o n tro l F e e d and feeds incorporated with 5, 10 and 15% o f dried fermented c a b b a g c w a ste .
Plate V: Solid S tate F'crm entation o f c ab b a g c waste in pctri plate and in conical flask with D. c o a g u la n s,
Plate VI: Solid State F e rm e n ta tio n o f C otto n seed cake in petri plate and in conical llask w ith B. c o a g u la n s.
Plate VII: Solid S ta te F e rm e n ta tio n o f G ingelly oilcake in pctri plate and in conical flask w ith B. coagulans.
Plate VIII : D ried c a b b a g c w aste after b e in g subjected to ferm entation with B. coagulans.
Plate IX : D ried C o tto n seed cakc after being subjected to fermentation with B. caagulans
Plate X ; D ried G in g c lly oilcakc after being subjected to fcrnientatio/i w ith B. coagulans
Plate XI: Solid state ferm en tatio n o f cab b ag e waste, cotton seed cake and gingclly oil cake in petri p la te s w ith B ea u veria sp..
Pkitc Xll; Solid state ferm entation o f cab b ag e waste, in conical llasks with B eauveria s p ..
Plate XIII: Solid stale ferm entation o f gingclly oil cakc and cotton seed cake in conical flasks.
Plate XIV: f h e e x p e rim e n ta l feeds c o m p o u n d e d incorporating with 5, 10 and 15 % o f dried c a b b a g e w aste ferm ented using Bacillus coagulans.
Plate XV: 'f h e e x p e rim e n ta l feeds c o m p o u n d e d incorporating with 5. 10 and 15 % o f dried c ab b a g e w a s te ferm ented using B eauveria sp..
Ill
Table 1. Proximate composition of some selected feed ingredients of plant origin in India.
Table 2. Some typical SSF processes used for the production of protein/ animal feed.
Table 3. Percentage composition o f the ingredients used in the control feed formulation.
Table 4. Percentage composition of the experimental feeds prepared using varying concentrations o f dried powdered cabbage waste.
Table 5. Proximate chemical composition of dried unfermented cabbage waste, gingelly oilcake and cottonseed cake.
Table 6. Proximate chemical composition of other feed ingredients used in the compounded feed base.
Table 7. Physical characteristics of the control and experimental feeds compounded utilizing dried, unfermented cabbage waste.
Table 8. Proximate chemical composition o f control and experimental feeds compounded utilizing varying concentrations of dried unfermented cabbage waste.
Table 9.Proximate chemical composition o f cabbage waste, cottonseed cake and gingelly oilcake after fermentation with B. coagiilans.
Table 10. Proximate chemical composition o f cabbage waste, cottonseed cake and gingelly oilcake after fermentation with Beauveria sp. fungi.
Table II. Changes in percentage o f some biochemical parameters upon Solid State Fermentation o f cabbage waste, cottonseed cake and gingelly oilcake.
Table 12. Physical characteristics o f the control and experimental feeds compounded utilizing cabbage waste fermented with B. coagulans.
Table 13.Proximate chemical composition o f the control and experimental feeds compounded utilizing cabbage waste fermented with B. coagulans.
Table 14. Physical characteristics o f the control and experimental feeds compounded utilizing cabbage waste fermented with Beauveria^p. fungi.
Table 15. Proximate chemical composition o f the control and experimental feeds compounded utilizing cabbage waste fermented with Beauvcria sp. fungi.
A C K N O W L E G E I M E N T
I w o u l d lik e to c o n v e y m y d e e p e s t r e g a rd s to m y C h a i r m a n a n d M a jo r A d v i s o r D r. ( M r s . ) M a n p a l S h r id a r , S c ie n tis t, C .M .F .R .I w h o h a s b een s u p p o r t i n g m e a l t h r o u g h m y w o r k a n d r e n d e r e d ti m e ly h e l p a n d s u g g e s ti o n s to m e in c o m p l e t i o n o f t h i s re s e a rc h . W o r d s c a n n o t e x p r e s s m y g ra titu d e t o w a r d s h e r.
1 w o u l d a l s o lik e to c o n v e y m y r e g a r d s to m y c o - g u i d e D r.R .P a u l R aj, C o - C h a i r m a n , A d v i s o r y c o m m i t t e e a n d H e a d , P . G .P .M , C .M .F .R .I fo r c r e a tin g a s u i t a b l e e n v i r o n m e n t f o r th e s u c c e s s fu l c o m p l e t i o n o f t h e w o r k and t i m e l y m o d i f i c a t i o n s th a t w e r e n e c e s s a r y fo r p r o d u c i n g a g o o d q u a l i t y w o rk .
Sri. P . V i j a y a g o p a l , a m e m b e r o f m y a d v i s o r y c o m m i t t e e w h o h a s b e e n p r o v i d i n g t h e n e c e s s a r y h e lp a n d s u p p o r t e s s e n tia l fo r m e. I s in c e r e ly th a n k h i m f o r p r o v i d i n g th e c o m p u t e r fa c ilitie s a n d g u i d a n c e r e q u ir e d in c o m p l e t i o n o f m y w o r k .
I e x p r e s s m y g r a t i t u d e to D r. M . S r in a t h fo r b e i n g a m e m b e r o f m y a d v is o r y c o m m i t t e e a n d h e l p i n g in t i m e s o f n e c e s s it y a n d D r . S u d h a k a r R a o for s u p p o r t i n g m e in t i m e s to g r i e f a n d m is f o r t u n e .
1 w o u l d a l s o l i k e to t h a n k D r . V . C h a n d r i k a , D r.K .S u n il K u m a r M o h a m m e d , D r. N o b e l a n d A b o o b a k a r a n d all th e o t h e r s c ie n tis ts w h o h a d b e e n helpJlil in o n e o r t h e o t h e r w a y .
I w o u l d lik e to th a n k M r . N a n d a k u m a r fo r te c h n ic a l a ss is ta n c e , M r . N . R u d h r a m u r t h y , t e c h n ic a l a s s is ta n t , D F D , M r. M .P .P a u lto n and S a ty a se e la n a n d J o y a n d G ir is h fo r t h e i r h e lp in p r o v i d i n g s elfle ss a s s is ta n c e w h e n n e e d e d .
1 a m e n d o w e d t o M r. M .B . S e y n u d e e n a n d M r. P.P. P a v ith r a n for p r o v id i n g m e w i t h n e c e s s a r y c o m p u t e r fa c ilitie s a n d h e l p i n g m e ta k i n g the p rin to u ts.
1 a m g r e a t l y o b l i g e d to M r. P .S ita p a ti B a b u w h o w a s v ery frie n d ly and h e lp f u l in c o m p l e t i o n o f th is w o rk . I w o u ld a lso like to th a n k K ira n bab u , a n d all o f t h e p e o p l e w h o w e r e r e s p o n s ib le fo r th e c o m p le tio n o f th is w ork. T h a n k i n g y o u o n e a n d all.
I w o u l d lik e t o e x p r e s s m y t h a n k s to In d ia n C o u n c i l o f A g ric u ltu ra l R e s e a r c h , N e w D e l h i f o r p r o v i d i n g fe llo w s h ip .
I. IN T R O D U C TIO N
In the recent years, there has been a rapid developmenl in the field o f fisheries especially with reference to aquaculture. India has vast fisheries resources with a total EEZ (Exclusive Economic Zone) of 2.02 million square kilometers and a coastline of 8047 kilometers. The total riverine length is above 29,000 square kilometers, fhe lentic water bodies include lakes, ponds, tanks, reservoirs, bheels, bans, jheels, ctc/vhich arc estimated to be nearly 1.29 lakh hectares in area. The rivers upon joining the sea form large areas o f brackish water estuaries. The area suitable for braekishwater fish farming is around 2 million hectares. These form a tremendous resource for fisheries in the country. The marine environment produces around 2.6 million tonnes o f fish, comprised o f different variety o f fishes, crustaceans and mollusks. The inland water resources produce around 2.3 million tonne o f Hsh. Eventhough the current inland production lags bciiind liic marine production there has been a tremendous increase in (he annual growth rate primarily due (o aquaculturc.
In the early 90's shrimp farming changcd the aquaculturc scenario with vast strclches o f coastal lands being converted to shrimp farms generating valuable foreign exchange for the country. Simultaneously freshwater prawn farming using the species Macrobrachium rosenhergii was also initiated.
In aquaculture more than half the investment comprising 50 to 70 % of the total operating costs goes into feeds as they contribute an essential factor for enhancing f'lsh production. With the intensification o f culture activities more emphasis is diverted to fish nutrition and compounded feeds in particular.
Though fish are probably the most efficient feed converters, there exists a dearth in the availability o f food supply for cultured fish, 'fhereibrc, there is necessity to look for newer or non-conventional feed ingredients to cover this deficit. Fish nutritionists have long tried to use less expensive plant protein sources to partially or totally replace fishmeal as well as shrimp meal. O f all the plant protein ingredients, soybean meai is
considered to be the m ost nutritious and is currently used as the major protein source in many fish diets. However, growth tended to be reduced in fish fed with diets in which soybean meal completely replaced the fishmeal, which could be attributed to the anti- nutritional tactors present as well as to the lower assimilation efficiencies o f plant protein in comparison to animal proteins. It is therefore, important to identify alternative protein sources to reduce or stabilize the cost o f fish production, as ultimately econom ic factors are the ones, which determine the level o f inclusion.
Non conventional ingredients are ingredients that are capable of partially or completely substituting fishmeal. These have been in use since traditional aquaculture in Asia. It has been found that these feedstuffs can be used as a substitute for tlshmeal as they are no more abundant than fishmeal but are least expensive. Fishmeal has a well-balanced amino acid profile along with essential fatty acids. All foodstuffs need not have the same amounts o f amino acids in fishmeal but they usually exceed the levels found in som e am ino acids. Example is com gluten, which is low in lysine and tr>'ptophan but rich in other amino acids. These ingredients are considered to be satisfactory as feed constituents as they usually have the necessary levels of nutrients that are required for a given species. Ex: Herring meal has been known to fully meet the amino acid requirements o f trout.
Analysis o f m ost conventional feed ingredients however, indicates that no single ingredient can be used as a complete alternative or substitute for fishmeal. Also there exist some problems with these ingredients. Unconventional ingredients from plant origin have low protein and high carbohydrate and fiber content in addition to anti- nutritiona! factors and toxins. The commonly used non-conventional feed ingredients include krill meal, single cell proteins, rape seed meal, sunflower meal, poultry byproducts, feather meal, fly larvae, etc.
According to Akiyama (1991) the commercialization o f aquaculture is growing, thereby increasing the demand for aquaculture feeds. Traditionally, these feeds have been based on animal protein. However, due to cost and availability considerations, it is
inevitable that more plant protein supplements be utilized in feeds. Plant protein supplements are a more cost-effective source o f nutrients as compared to animal protein supplements.
I he advantages o f femientation have been known over the ages as a means of bioconversion and protein enrichment o f food and feed ingredients. It is also increasingly evident that the development o f low cost, high quality protein foodstuffs is crucial for the tuture success of the aquaculture industry. Solid state fermentation is a novel technology by means of which cheap ingredients o f lesser nutritive value can effecti\ely be c o n \’eried into nutritionally rich and easily digestible aquafeeds (Nigam and Singh. 1996).
Cabbage {Brassicu olaracea var. capiici(a) is available in the local vegetable markets throughout the year. The outer green leaves are usually discarded as waste and only the inner com pact head is utilized. The waste is available in bulk in most market and ils incorporation, as a non-conventional ingredient in shrimp feeds would therefore be a hicrativL* proposition. As gingelly oilcake and cotton seed oilcake are also commonly used in shrimp feeds it would be most pertinent to carry out further enrichment o f these substrates as well along with cabbage waste in order to increase their nutrili\e value and digestibility. The present study was therefore proposed with the following main objectives:
1. Proximate com position analysis o f cabbage waste, gingelly oilcake and cotton seed oil cake.
2- Fermentation o f cabbage waste, gingelly oilcake and cottonseed oilcake individually using both bacteria and fungi.
3. Formulation o f feeds with fermented cabbage waste in permutation combination with other conventional feed ingredients.
4. Determination o f the physico-chemical characteristics o f these feeds using standard melhodoloeies.
2. REVIEW OF LITERATURE
Nonr conventional Feed I n g r e d i e n t s
On a purely nutritional basis it has been repeatedly shown that the best food or feed ingredient in terms o f nutritional composition palatability, growth and feed conversion efficiency is fishmeal. However, its high cost and competition for use as human food as well as other terrestrial farmed animals makes its availabihty scarce.
Commercial aquaculture feeds for shrimp generally contain 25-45% of crude protein because shrimp require such high dietary levels. Consequently only high protein oil seed residues have been used for compounding shrimp feeds (New, 1976).
Feedstuffs o f vegetable origin as a whole are lower in protein when compared to those of animal origin. Nevertheless among all plant protein sources tested for most crustaceans, soyabean meal has been found to be the most superior on account of its high protein content and essential amino acid profile (Kanazawa, 1995; Akiy&ma,
1988).
In order to reduce the escalating cost of aquafeed and make aquaculture sustainable in the long run ^intensive research is being focussed on altemative and more sustainable protein sources for use within compounded aquafeeds (Tacon, 1993). The utility o f plant protein as partial replacement for the more expensive animal protein fractions has been examined but results show great variations in the degree of success, which inordinately depend on the species and types of ingredients used. Lim and Dominy (1989) and Tacon (1993) reviewed the utilization o f plant proteins in formulated feeds for warm water fishes. A compendium on the potential use of plant protein in marine fish feeds was given by New (1989). A large number of plant ingredients have been used in animal feeds and fish feeds but relatively few in shrimp
feeds. The extent o f plant protein utilization is normally influenced by the availability, cost, acceptability, ease o f processing, the nutritive value and presence of toxins or anti- nutritional factors (Akiyama.1991).
Tacon (1993) reviewed utilization o f oil seeds and their by-products for warmwater fish species. Peanut meal proved beneficial as a source o f plant protein for penaeid prawns and its use as oilcake was reported by All (1994). Raman et a i, (1982) successfully tested artificial feeds prepared from Bengal gram husk, gingelly oilcake, bajra along with fishm eal and prawn waste meal and advocated their use for reducing feed cost. Efficiency o f cottonseed meal was examined in the diets o f Macrobrachium rosenhergii and P enaeus indicus (Aquacop, 1976; Aii, 1994).
K eem biyahetty and DeSilva (1993) reported the replacement o f 20-33%
fishmeal by cow p e a and black gram in diets for Oreochromis niloticus fingeriings.
Wheal flour, wheat gluten and rice bran were recommended for use in feeds o f Penaeus indicus by Galgani et a t , (1989) and Paul Raj (1990).
With pressure mounting to increase the production o f aquaculture feeds, tapping of non-conventional and alternate feed resources becomes pertinent. A variety o f non- conventional feed resources such as single cell proteins (SCP), leaf meals, aquatic plants like seaweeds, water hyacinth. Coontail, aquatic fern, duckweed, water lettuce were identified for incorporation into fish and shellfish feeds {^ew, 1976; Tacon, 1990;
Tacon, 1993). C assav a or Tapioca meal {Manihot esculentd) alfalfa or clover, leaf protein concentrate and coffee were successfully incorporated in feeds o f warmwater ftshes (Ng& wee, 1989;Viola et a i , 1988; Jia et a l , 1991 Chow et a i, 1983; Ogino et a l, 1978 and C hristensen 1981). Syslo and Hughes (1981) and Harpazand Schmalbach (1986) demonstrated the benefits o f supplementing dry artificial diets with fresh leaves for lobster and M acrobrachium rosenbergii respectively. Catacutan (1993) showed that aquatic macrophyte N aja graminae and Ruppia martima were consumed by Penaeus monodon with high assimilation efficiencies o f between 40-76%. Aquatic macrophytes were used as live sources for ^iacrobrachium rosenbergii (Stern et al.. 1976). Use of
sea~grass for mud crab was evaluated by Raman el a}.. {1982). These studies showed that live plant organic matter was well utilized by crustaceans.
Solid State F e r m e n ta t io n :
If the aquaculture sector is to reduce its dependence upon fishmeal and farmers are to bring dow'n feed and production costs and improve farm profitability it is right time that modern technology be adopted. O f the \s'ide jvariety ofi feed ingredients available in India for production o f aquafeed^(N ew et a l, 3’993) most are reported to be of too poor quality to produce high quality aquafeeds, especially for shrimp. The proximate composition o f various commonly used conventional and non-conventional plant ingredients is given in Table, 1.The noticeable variability is due to a number of factors including the m ethod o f handling and processing and the nutritional status o f the environment in w^hich they were grown, in addition to variations in analytical methodologies.
Solid State fermentation (SSF) has gained importance in the recent past due to its several advantages over submerged fermentation especially for enrichment of protein o f agricultural wastes and sub products. The SSF technology has the advantage of direct utilization o f none or very few pretreated solid substrates under aerobic conditions to produce microbial Biomass products (MBP), which contain a mixture o f unused substrates, cell substances o f the microbes and externalized metabolites (Nigam and Singh, 1996).
Some typical substrates used for SSF processes are given in Table 2 but most o f these were employed mainly for production o f enzyme protein and animal feed.
Cassava (Manihot esculenia Craniz) has been subjected to SSF for protein enrichment by a wide variety o f researches using filamentous fungi using Aspergillus niger (Senez 1979; Raimbault ef a l , 1985) and Rhizopus genus (Daubresse et a l, 1987; Mitchell et
6
al, 1988 and Soccol et a l, 1993). The effect of substituting corn with fermented cassava in broiler rations was investigated by Varghese el a l, (1996), while ManiJal et al.. 1987 used A spergillus niger for utilizing starch factoo' waste by SSF. Protein enrichment o f cassava flour and starch factory wastes using the fungus Trichoderma pseudokongii Rifai in a solid state fermentation process for incorporation in cattle feeds and broiler rations was developed by Balgalagoplan and Padmaja (1988) and Padmaja and Balagopalan (1990). Biotransformation o f a number o f crop residues into animal feeds was also achieved by SSF.
Though microbially fermented fish silages and single cell proteins are being used in fish and shellfish diets the advent o f solid state fermentation in aquatic diets is comparatively new with very little research having been carried out to date.
The effect o f fermentation on the nutritive value o f seasame seed meal in the diets for rohu {Laheo rohiia) fmgerlings was reported by Mukhopadhay and Ray (1999). Migher feed effects were recorded in Moina macrocopa when cultivated on fermented grass pulps (Yang, 1995). Shimeno et a l, (1993) determined the effects on growth, feed conversion and body composition o f fermented defatted soyabean meal (SBM) either u ith A sp erg illu s oryzae or Eurotium repens in single moist pelJet diet for juvenile yellowlail S erio la quinqueradiata. Hossain et a l, (1988) reared red sea bream Chrysophrys m ajor on highly o.xidized scrap meal fermented by a group o f microbes and compared their grow th and feed efficiency to fish reared on non fermented and defatted scrap meals and white fish meal. Fermented vegetable and kitchen wastes were recommended as feeds for not only zoea but also mysis and upto certain points in the post lar\'al stages o f P enaues monodon, w'hen diatoms or brine shrimp nauplii were lacking or in short supply.
Sridhar and Chandrashekar (1996) employed SSF technology for fermentation of groundnut oilcake and wheat bran for incorporating into shrimp feeds. The fermented material at varying concentrations was incorporated into feeds and its effect on growth and feed conversion efficiency o f Penaeus indicus post larvae studied.
7
1 he studies carried out so far are scarce but offer immense scope for research to prove beyond doubt, that solid slate fermentation, which is simple and economic, is the appropriate technology for the futuristic aquafeed industry. Though not complete in itself the present investigation is also an attempt towards this direction.
T a b l e 1 .P r o x i m a t e c o m p o s i t i o n o f s o m e s e l e c t e d f e e d i n g r e d i e n t s o f p l a n t o r i g i n in I n d ia .
i w B l p g f t q i c n t ^ ^
Rice Polish 12.6 14.5 17.3 7.5 - -
Rice Polish 10.0 12.2 16.0 9.0 6.0 46.8
Rice Polish 8.4 11.4 15.3 n . o 12.9 41.0
Rice. Broken 10.0 12.0 4.2 5.3 3.1 65.4
Rice Bran 10.1 12.6 11.3 19.3 10.2 36.5
Rice Bran 7.8 7.8 6.1 4.4 20.5 43.4
Rice Bran 8.4 2.9 5.0 18.0 20.3 38.4
Rice Bran 8.7 9.4 4.7 13.5 31.4 32.3
Defatted Rice Bran 7.2 12.1 1.3 15.2 23.8 40.4
Wheat Bran 12.3 15.8 4.3 8.7 - -
Wheat Bran lO.O 13.5 2.6 12.2 3.0 58.7
Wheat Bran 13.0 8.2 6.6 33.5 4.2 34.5
Wheat Bran 9.3 12.6 7.5 11.9 4.2 54.5
Wheat Broken 9,0 11.5 1.9 4.0 0.2 73.4
Wheat flour 12.6 14.5 3.7 2.7 2.3 64.2
GNOC 7.8 28.6 13.8 7.5 13.4 28.9
GNOC 6.0 37.7 11.5 13.2 7.3 24.3
GNOC 10.0 42.0 7.3 13.0 2.5 25.2
GNOC 8.3 46.6 7.7 6.5 7.7 23.2
GNOC 7.1 35.8 8.5 8.2 10.5 29.9
Groundnut extract 7.0 48.0 2.0 11.2 2.7 29.1
Sunflower Extract 8.0 31.0 2.1 18.4 1.5 39.0
Sunflower Extract 10.2 30.1 2.9 24.7 6.5 25.6
Palm kernel cake 8.9 12.2 4.9 25.6 2.6 45.8
Soyabean meal n . 8 46.3 1.3 5.0 -
Soyabean meal Soyabean meal Soy sauce waste
3.0 10.0 12.0
58.6 46.0 13.5
1.4 0.9 8.2
0.4 7.3
5.8 _
5.3 0.6 5.3
31.3 35.2 55.2
Rapeseed cake 11.0 35.9 0.9 13.2 6.9 32.1
Sal seed cake 8.6 8.2 2.9 1.7 10.2 68.4
Sesame cake 8.3 41.9 9.2 6.2 14.8 19.6
Sesame cake lO.O 29.0 12.9 18.3 lO.O 19.8
Sesame cake 10.0 42.7 6.9 5.7 12.9 21.8
Mustard cake 8.5 30.8 9.3 6.2 10.3 34.9
Mustard cake 9.2 23.6 9.6 6.3 10.4 40.9
Cotton seedcake 7.0 37.0 6.7 13.0 1.0 35.3
Cotton seedcake 8.2 42.7 1.0 12.6 8.2 27.3
Gingelly cake 9.0 34.0 7.8 7.9 3.1 38.2
Gingelly extract 7.0 40.0 2.0 9.7 2.9 38.4
Niger ext. 7.0 35.0 2.0 19.0 3.5 33.5
Copra cake 12.0 22.0 6.5 12,2 5.2 42.1
Copra cake 8.4 20.3 11.4 16.2 6.2 37.5
Copra cake - 22.0 6.0 12.0 2.1 -
Tobacco seed ext. 7.7 30.6 0.3 - 13.7 47.7
Maize meal 13.5 9.5 4.0 4.0 1.5 67.5
Maize 10.4 4.6 7.8 3.5 1.0 72.7
Sorghum 10.0 9.0 2.8 3.0 0.1 75.1
Spirulina 8.7 50.5 I.O 2.1 II.O 26.7
Tapioca Flour 11.5 3.1 2.3 2.0 2.3 78.8
Tapioca Flour 8.0 1.8 1.3 1.8 0.2 86.9
Coffee pulp 12.3 14.0 1.2 20.8 8.2 43.5
Colocasia meal 5.8 24.6 4.5 8.2 9.9 47.0
Eichornia meal 3.3 19.5 2.3 18.3 9.3 47.3
Pistia meal 4.9 19.5 1.3 11.7 25.6 37.0
Leucaena meal 11.8 33.1 4.7 9.0 7.2 34.2
Mulberry leaf, dry 8.9 27.2 2.4 I I . 5 8.1 41.4
Salvinia meal 2.6 16.2 l . l 18.5 22.0 39.6
Sources: Michael B .N ew , Albert G.J. Tacon and Imre Cavas- Farm - made aquafeeds.
T a b l e 2 . S o m e T y p i c a l S S F P r o c e s s e s U s e d F o r t h e P r o d u c t i o n o f P r o t e i n / A n i m a l F e e d
SI. N o ' ‘ StJ B S T R A T E S--- MICROORGANISMS - r ■""‘"YEAR '
1. Canola meal Aspergillus carbonarius Alachel & Duvnja (1995)
2. Carob pods Aspergillus niger Smail et a l (1995)
Apple pomace Candida utilis
Kloeckera apiculata
Rahmat et al. (1995)
4. :*Coffee pulp Penicillium verrucosum Roussos et al. (1994)
5- i Sugarcane bagasse Chaetomium cellulolyticum Brabo et al. (1994)
6 Sugarbeet pulp &
molasses
Fusarium oxysporum Chaetomium cellulolyticum Trichoderma reesei
Trichoderma viride Saccharomyces cerevisae
Nigam (1994)
7 Apple pomace Candida utilis
Candida tropicalis T.viride
A.niger
Bhalla and Joshi (1994)
8 (Grange peels. Grape stalks
Pleurotus ostreatus Agrocybe aegirata Armillarialla mellea
Nicolini et al. (1993)
1 9 Cassava Rliizopus arrhizus Soccal et al. (1993)
10 Oil Palm wastes C.cinereus
P.sajor caju
Kume et al. (1993)
Cassava A.oryzae Zvauya and Muzondo
(1993) 12
1
Cotton stalks. Perlite P.ostreatus Ph.chrsosporium
Kerem and Hadar (1993)
13 Wheat straw P.ostreatus Tripati and Yadav (1992)
14
i
! Grape pomace. Corn Chaetomium cellulolyticum+ Girujie et al.
! stover Candida utilis (1992)
15 Rice straw. Maize stover Cyathus stercoreaus Dichotomus squalens Ph.chrysosporium
Karunanandaa (1992)
16 Sugarcane bagasse Polyporus sps Nigam (1990)
17
1
1 Sugarbeet pulp i
T.reesei, Fusarium oxysporum
Nigam et al. (1990) ' 18 Sugarcane bagasse Mixed cultures
(poluporus. Pleurotus, Trichoderma)
Nigam (1989)
19 Corn cob A.niger Singh et al. (1989)
20 Siarchy raw materials A.oryzae, A.niger, A.awamori
Czajkowska et al. (1988)
21 Sugarbeet pulp T.viride Durand et al. (1988)
22 Sugarbeel pulp T.reesei
Fusarium oxysporum
Nigam etal. (1988)
23 Wheat straw Coprinus sps Yadav(1988)
24 Sugarbeet pulp Thermophilic fungi Grajek(1988)
25 Cassava Rhizopus oryzae Daubresse e ta l. (1987)
26 Saccharum munja
residues
Piuerotus sps Gujral et al. (1987)
27 Straw Candida utilis Han (1987)
28 Cassava S.pulverrientum Smith et al. (1986)
29 Banana wastes A.niger Baldensperger etal(1985)
30 Dried citrus peel A.niger Rodriquez e ta l (1985)
31 Wheat straw T.reesei
Ch.cellulolyticum C.utilis
Abdullah et al. (1985)
32 Sugarcane bagasse Polyporus sps Nigani e ta l. (1989)
'y Wheal straw T.reesei
Endomycopsis fibuliger
Laukevics et al. (1984)
34 1-odder beets Saccharomyces cerevisae Gibbon et al. (1984)
35 Pulpmill waste Ch.cellulolyticum Tautorus et al. (1983)
36 Soya bean Rhizopus oligosporus Rath bun et al.(1980)
37 Cassava T.reesei+yeast Opoku et al. (1980)
38 Starcli substrates Various cultures Senez et al, (1979)
39 Alfaalfa A.terreus Bajracharya e ta l.(1979)
4U Straw, corn sto\'er Ch.cellulolyticum Moo-Young et al. (1976)
4! Rye grass Cellulomonas, A.faecalis
T.reesei. A.pullulans
Yu et al. (1976)
42 R> e grass C.utilis Han. &
Anderson. (1975)
43 Newsprint Sporotrichum themophile Barnes (1972)
Sourcc: Uludai:, S.. P ro k o p , A. and T a n n e r R.D (1996).
Plate I: Fresh cabbage w aste and cooked, dried and powdered cabbage waste.
3. M A T E R I A L S A N D M E T H O D S
T h ree n o n -c o n v e n tio n a l feed ingredients viz. cabbage waste, gingelly oilcake and c o tto n s e e d o ilc a k e p ro cu red fro m the local m ark et were analyzed for their p ro x im a te c o m p o sitio n . Feeds w e re c o m p o u n d e d with v a ry in g c o n centrations o t c a b b a g e w aste a n d th e ir physical and physico-chem ical characteristics d e te r m in e d . T h ese three ingredients w e re subjected to Solid State F e rm en tatio n ( S S F ) u s i n g b o th a bacillu s {Bacillus coagulans) and a fungus (B ea u veria s p .) a n d th e ch an g e s in c h em ical com p o sitio n were determ ined.
Feeds o f v a r y i n g c o n c e n tra tio n s w e re again form ulated w ith the ferm ented cabbage w a ste a n d th e p h y sica l and p h y sico -ch em ical characteristics c o m p a red to those o f th e u n f e r m e n t e d m aterial.
Selection of Ingredients
R aw m a te ria ls fo r feed fo rm ulation i.e. fishmeal, soyabean meal, shrim p meal, w heat flour, g ro u n d n u t oilcake, gingelly oilcake, cottonseed cake w ere procured from th e lo c a l m ark et. T h e y w e re sorted o f f extran eo u s material, oven- dried, pow'dered in a p u lv e ris e r and sie v e d through a sieve o f 250 m icrons m esh size. C ab b a g e w a s t e w a s co llected and transp o rted to the laboratory. T h e leaves were rinsed s e v e ra l t im e s in tap w a ter a n d finally in double distilled water. After draining for o n e h o u r th e y w^ere dried in an oven at 60 ± 5 C and p o w d e re d as m entioned earlie r (P la te 1). P ro x im a te com p o sitio n analyses o f ingredients w as perform ed fo r w h i c h c h e m ic a ls w e re p u rc h a se d from stan d a rd chem ical firm s o f national and in te rn a tio n a l repute.
Feed f o r m u l a t i o n :
The feed ingredients and other additives like oil, binder, vitamin premix, mineral mix were w eighed according to the standard feed formulation given by New* 198?'
T a b l e 3 . P e r c e n t a g e C o m p o s iti o n o f the I n g r e d ie n ts used in the C o n tr o l F e e d F o r m u la t io n
. . .
Cabbage waste 0.0
Fish meal 15.0
Soya bean meal 36.0
Shrimp Meal 10.0
Wheat flour 18.0
Ground Nut Oil Cake 10.0
Cod liver Oil 2.0
Vegetable oil 2.0
Gelatin 4.0
Vitamin mixture 0.5
Coated vitamin C 0.5
Mineral Mixture 1.0
Di-Calcium Phosphate 1.0
• Standard USP XVII mixture procured from SISCO laboratories.
Preparation o f control feedi
Fish meal, soyabean, meal, shrimp meal, wheat flour, groundnut oilcake were weighed individually as per the specifications mentioned in Table 3 and
13
mixed h o m o g e n e o u s ly . G e latin w as d isso lv e d in sufficient w ater and h eated till it dissolved c o m p le te ly . W h e a t flour w a s th e n added and gelatinized. T h e m ixture was cooled a n d th e o th e r ingredients and v itam in p re m ix and m ineral p rem ix, di
calcium p h o s p h a te c o a t e d v itam in C a n d m ix e d h o m ogeneously to obtain a dough. This d o u g h w a s p a lle tiz ed into n o o d le s usin g a kitchen m incer fitted w ith a 2 -m m die. P ellets w e r e initially su n d rie d and then dried in an oven at 60'" C ± 5'’ C for 12 hrs to le ss t h a n 10% m o istu re content. T h e y w ere m anually b ro k e n into sm aller bits a n d s to re d in a plastic c o n tain er at ro o m temperature. T h e se dried pellets s e r v e d a s th e control feed.
Preparation o f e x p e r i m e n t a l c a b b a g e w a st e feeds:
T h e e x p e r i m e n t a l diets w e re p re p a re d as p er the above form ulation (T able 4) except that fish a n d sh rim p m eal w e re equally replaced with 5%, 10% and 15% dried c a b b a g e w a s t e and these feeds w e re designated as CWI, C W II and CWIII resp ectiv ely .
Table 4: P e r c e n t a g e c o m p o sitio n o f the E xperim ental Feeds prepared using v a r y i n g c o n c e n tr a tio n s o f dried pow dered ca b b a g e waste.
IN G R E D IE N T S '/o
E X P E R IM E N T A L FEEDS
C W I C W II C W III
Cabbage waste 5.0 10.0 15.0
Fish meal 12.5 10.0 7.5
Soya bean meal 36.0 36.0 36.0
Shrimp Meal 7.5 5.0 2.5
Wheat tlour 18.0 18.0 18.0
Ground Nut Oil C'ake 10.0 10.0 10.0
Cod liver Oil 2.0 2.0 2.0
Vegetable oil 2.0 2.0 2.0
Gelatin 4.0 4.0 4.0
Vitamin mixture 0.5 0.5 0.5
Coated vitamin C 0.5 0.5 0.5
Mineral Mixture* 1.0 1.0 1.0
Di-Calcium
Phosphate 1.0 1.0 1.0
Total 100.0 100.0 100.0
* Standard U S P X V II m ix tu re p ro cu red Irom SISC O laboratories.
Preparation o f f e r m e n t e d ca bba g e w a ste feeds:
A set o f th re e feeds w e re c o m p o u n d e d by incorporating 5, 10 and 15% o f cabbage w a ste fe rm e n te d using B .c o a g n la n s and designated as C W IV , C W V , CWVI re sp ec tiv e ly and a set o f th ree feeds each form ulated by incorporating 5,10 and 15% c a b b a g e w aste ferm ented using B eauveria sp. and designated as C W V ll. C W V ll l a n d C W I X respectively. T h e diet devoid o f tcrmcnted material designated as C s e r v e d as control. T h e percentage incorporation o f other feed
ingredients used in th e feed b ase w a s as given in T a b le .3. The feeds w ere prepared in th e s a m e m e th o d as d e sc rib e d earlier. T h ey w e re dried to less than ten- p ercen t m o is tu re c o n te n t and sto re d in airtight plastic containers at ro o m tem perature till fu rth e r analy sis.
Physical ev a lu a tio n o f t h e feeds:
T h e p h y sica l a p p e a r a n c e o f th e feeds viz. color, shape, size and pellet d iam eter w e re re c o rd e d .
H v d r o sta b ilitv tests o f th e feeds:
W ater s tab ility o f th e con tro l feed w as d eterm in ed by the m e th o d o f J ay aram and S h etly (1 9 8 1 ) w ith m in o r m odifications. A p p ro x im ately 5 g o f diet w as w e ig h ed , in trip lic a te and tra n sfe rre d 4 ’' X 4” p o u c h es m a d e o f boltin g silk.
T hese w ere im m e r s e d in 25 liters o f sea w'ater (28ppt salinity and 28 ± 2" C w ater te m p e ra tu re ) in plastic tubs. P o u ch es w ere re m o v e d from w a ter at 0 .5 ,K 2 ,3 an d 4 h o u rs re sp ec tiv e ly a n d rinsed w ith distilled w ater to re m o v e adhering salts. T h e c o n te n ts w e re tra n sfe rre d to p re -w e ig h e d petridishes and dried in an o v e n at 80 ±5 C and the re su lta n t loss in dry m atter was calculated.
Plate II: Fresh culture o f Bacillus coagulans used for solid state fermentation o f cabbage w aste (C W ), Cottonseed cake (C SC ) and Gingelly oilcake (GOC).
Plate III: Dried and powdered cottonseed cake and gingelly oilcake before inoculation with bacteria and flingi for fermentation.
SOLID S T A T E F E R M E N T A T I O N :
Solid State F e rm e n ta tio n o f ingredients w as p re fo rm ed by follo w in g the method o t R a m e s h a n d L o n s a n e (1987). N e c e ss a ry changes w e re how ever, m ad e in the m e th o d o lo g y a n d m e d iu m c o m p o sitio n after optim ization o f p ro cess parameters.
M ic r o -o r g a n i s m s :
Pure c u ltu re s o f B a c illu s c o a g u la n s (Plate II) and fungi (B eauveria sp.) were p ro c u re d f r o m th e m ic ro b io lo g y section o f th e D e p artm en t o f B iotechnology, C o c h i n U n iv e rsity o f S cience and Technology. T h e y w e re maintained as p u re c u ltu re s by s u b -c u ltu rin g every w eek on nutrient agar slants tor baclcria a n d m y c o l o g i c a l a g a r c o n ta in in g streptom ycin for fungi.
Preparafion o f s o lid s u b s t r a tu m :
C ab b a g e w a s t e a l o n g with g in g e lly oilcake and cottonseed oilcake (Plate III) were d ried at 60 ± 5" C for 2 4 h rs in an oven. T h ey w ere p o w d ered in a pulveriser and s ie v e d th ro u g h a 2 0 0 -m ic ro n sieve to obtain uniform particle size.
The pi I o f eac h in g r e d ie n t w a s d e te rm in e d individually. M oisture content o f each substrate w as also e s t i m a t e d usin g I g o f th e sam p le and m oisture content o f the medium w as a d ju s t e d to a level v a ry in g b e tw ee n 50 and 60%. T h e solid substrates w ere d i s p e n s e d as 5g aliq u o ts in petriplates as also in 250 m l conical tlasks and adju sted to th e desired level o f m o istu re content w ith requisite a m o u n t o f physiological s a lin e ( 0 .8 5 % N a C i), a d ju sted to the o p tim u m pH (p H 8.0 to 12.0 for b acteria a n d fungi re sp ectiv ely ). T h e flasks and petriplates a lo n g w ith
their contents w e re a u to c la v e d at l 2 l ^ f o r 60min. and co o le d dow n to ro o m temperature.
Inoculum P r c p arntion'
i)Bacteria:
A lo o p fu l o t 24 h r old B .c o a g u la n s culture w as first grow n in 10ml nutrient b ro th to r 18 h rs at ro o m te m p e ra tu re (2 8 ± ‘’2C). 1 m l o f the above culture was tra n sfe rre d a s e p tic a lly to 50 m l n u trien t broth and incubated in a rotary shaker at I 5 0 r p m for 18hrs at ro o m tem perature. C ells w e re harvested by centrifugation at 10,000 rp m for 15 m in s at 4 ‘’ C T h e h arv ested cells w ere m ade up to 10ml u s in g sterile p h y s io lo g ic a l saline (0.85% N a C l) after repeated washings. T h is w a s u s e d as inoculum .
ii) Fungi:
20 m l o f s te rile p h y sio lo g ic al salin e containing 0 .1 % T w een 80 w as added to each fully s p o r u la te d (2 w e e k old) sla n t culture raised o n m ycological a g ar by means o f sterile p ip e tte . T h e spores w e r e scrapped u sin g a inoculating needle under strict ase p tic c o n d itio n s a n d th e sp o re suspension obtained w^as adjusted to the desired c o n c e n tr a tio n usin g sterile p h y sio lo g ical saline (0.85% N aCl).
Inoculation and i n c u b a tio n :
T h e p r e p a re d in o c u la w e re a d ju ste d to a concentration o f 2 m g dry cell equivalent in 1ml o f cell s u sp e n sio n o f bacteria and 2 m l o f fungal spore suspension p e r flask (in o c u lu m level s ele c te d rand o m ly ) for the fungi and added to sterilized m o ist m e d ia in flasks. T h e contents w e re m ix e d and incu b ated in
18
slanting p o sitio n at 35 " C a n d 28 C for b acteria and fungi respectively w ith 60- 70“o rclati\ e h u m id ity to r 48 to 72 hrs. P elriplates w ere in o cu lated and incubated s i m i l a r h . 1 h ey w e re p la c e d in the straig h t positions. A fte r ferm entation the contents w e re d rie d to a co n sta n t m o istu re level and a n aly zed for the various uulrient>.
Analytical M e th o d s :
M o istu re, a sh . c ru d e protein, c ru d e fat and fiber in feed ingredients, Icrmented s u b s tr a te s a n d feeds w e re d e te rm in e d by stan d a rd procedures (A O A C
1990) as giv en b e lo w .
M oisture c o n te n t:
M o istu r e c o n te n t in the feed ingredients, substrates and the c o m p o u n d e d feeds w ere d e t e r m i n e d g r a \ im etrically b y oven d ry in g the sam ples at 80 ° C till consecuti'.e s im ila r re a d in g s \sere o b tained. Percen tag es o f moisture in the sample read in g s w e re o b tained. P erc e n tag e o f m o istu re in the sam ples are calculated as tb llo w s;
Moisture c o n te n t % = w e ig h t o f fresh sam ple- w eig h t o f dry sam ple X 100 / weight o f fresh s a m p le
a\ sK content:
W'eiehed l\v\ s a m p le s o f th e feed ingredients and feed w'ere taken in duplicate silica c ru c ib le s, and ash ed in a m uffle furnace at 600 C for 6 hours.
Percentage o f ash w a s c a lcu la te d as follow's:
Ash % = w e is h t o f a sh X 100 / w e ig h t o f sam ple
Crude Protein:
Total n itro g en in the feed ingredients, substrates and feeds w as determined by th e "K je ld a h l’s method" and the result w as multiplied by a conversion factor o f 6.25 to give crude protein %.
Crude Fat:
Lipid c o n te n t o f feed sam ples w as determ ined by the Soxhlet’s Extraction M ethod usin g p e tro le u m ether (boiling p oint 60-80 C) as solvent.
Crude Fiber:
C rude F ib e r w a s determ ined as the fraction rem aining after digestion with standard so lution o f sulfuric acid (0.23N ) and Sodium H ydroxide (0.3 IN ) u n d er carefully co n tro lle d conditions.
R E S U L T S
P roxim ate c o m p o sitio n analysis o f cabbage w aste (C W ) was carried out with a v iew to assess its suitability for incorporation into shrim p feeds as a non- conventional feed ingredient. T h e p ro x im a te composition analysis o f tw o other conventional ingredients viz. cottonseed cak e (CSC) and gingelly oilcake (G O C ) was also c o n d u cted in ord er to fe rm e n t th e m and evaluate en hancem ent o f their nutritional v a lu e b y m icrobial enrichm ent, for incorporation into shrimp feeds.
Feeds w e re c o m p o u n d e d with 5,10 and 15% concentrations o f cabbage v/aste, both b efore a n d after fermentation. P e lle t characteristics, hydro^stability and nutritional c o m p o sitio n o f these c o m p o u n d e d feeds w as'evaluated.
I. P r o x i m a t e c o m p o s i t i o n o f C a b b a g e w a s t e ( C W ) C o t t o n s e e d c a k e ( C S C ) a n d G i n g e l l y o i l c a k e ( G O C )
Results on th e proxim ate co m p o sitio n o f cabbage w aste (C W ) are recorded in T a b le 5. Cooked and d ried cabbage w aste recorded a dry m atter content o f 9 5 .2 9 % a n d m oisture co n ten t o f 4.71%. T h e crude protein content averaged at 15% a n d th e fat content w as quite low recording an average valu e o f
1.50%. The cru d e fib re content o f 14.57% w as on the hig h e r side and ash content moderate at an a v e r a g e o f 11.46%. T h e carbohydrate content o f these w aste cabbage leaves w a s ab o v e 50%. T h e acid insoluble ash content o f 0 .0 9 % w as negligible.
C o tto n seedcake re c o rd e d 28 % protein, w hile gingelly oilcake recorded a h ig h er v a lu e at 34.4% as th e crude protein content. Values o f 5.22%
and 8.83% resp ectiv ely w ere obtained fo r the lipid content o f cottonseed cake
and gingelly oilcake. A com paratively high value o f 16.33% w as recorded for the fibre content o f c o tto n s e e d cake w hile a very low valu e o f 1.88% was re c o rd e d in the case o f g ingelly oilcake. The ash contents o f cottonseed cake and gingelly oilcake obtained in th e presen t study a v erag ed 6.02% for the former and 14.21%
for the latter. V a lu es o f 4 3 .8 8 % and 39.49%> w ere recorded for the nitrogen free extractives o f c o tto n s e e d cake and gingelly oilcake respectively. A low A IA value o f 0.29 w'as o b ta in e d in the case o f cottonseed cake w hile gingelly oilcake recorded a c o m p a rativ ely h ig h e r valu e o f 1.37% as the A IA .
Table S.Proximate chemical composition o f dried unfermented Cabbage waste fCW), Cottonseed cake ( C S O and Gingelly oilcake (GOC).
NUTRIENT INGREDIENT (%)*
CW CSC GOC
Dry matter 95.29 99.071 98.824
Moisture 4.71 0.929 1.176
Crude Protein 15.431 27.619 34.414
Ether extract 1.502 5.224 8.833
Crude fiber 14.568 16.333 1.875
Ash 11.455 6.019 14.209
Acid insoluble ash (AIA) 0.094 0.286 1.366
NFE** 52.335 43.876 39.493
* Values expressed on dry matter basis and average of three estimations carried out in triplicate.
** Nitrogen Free Extractives- calculated as (100-%crude protein +crude fat + ash + crude fiber + moisture)
The p ro x im ate com position o f fishmeal, shrimp meal, groundnut oilcake, soyabean meal and w h e a t flour used in com po u n d in g the feed was also evalu ated and results are re c o rd e d in Table 6. V alu es obtained in this study were in k e ep in g with values already reco rd ed in literature for these conventional feed ingredients.
Plate IV : T h e c o n t r o l F e e d a n d f e e d s in c o r p o ra te d w i t h 5, 10 an d 15% o f
dried ferm en ted c a b b a g e w a ste .
Tabic.6: Proximate chcmical comnosition of other feed ingredients used in the compounded feed base
NUTRIENT IN G R E D IE N T (%)*
PM SM GNOC SvM WF
Dry matter 99.271 99. 471 99.169 91.352 94.747
Moisture 0.729 0.529 0.831 8.648 5.253
Crude Protein 61.525 65.091 47.791 47.938 12.188
Ether extract 9.068 3.740 7.071 0.479 1.951
Crude fiber 0.399 3.154 1.714 5.229 1.268
Ash 23.46 17.369 6.520 6.858 1.45
Acid insoluble ash (A IA ) 0.517 1.244 1.509 ***ND
NFE** 4.818 10.115 36.264 30.849 77.889
* Values expressed on dry matter basis and a\'erage o f three estimations carried out in triplicate
** N itrogen I'ree Extractives- calculated as (100-%crude protein +crude fat+ ash + crude fibcr+m oisture)
***ND - N o t Detected.
II. P r o x i m a t e c o m p o s i t i o n o f c o n t r o l a n d e x p e r i m e n t a l f e e d s c o m p o u n d e d u s i n g c o o k e d , d r i e d u n f e r m e n t e d c a b b a g e w a s t e :
T h re e feeds w e r e f o r m u la te d in c o r p o r a tin g 5,10 a n d 15% o f c o o k e d , d rie d a n du n f e rm e n te d c a b b a g e w a s te in th e fe e d b a s e g iv e n in T a b le 4 (P late IV ) b y p a r tly re p la c in g fish m e a l a n d s h rim p m e al. A f te r c o m p o u n d in g the feeds w e r e d r ie d to less th a n 1 0 % m o i s t u r e a n d a n a ly s e d fo r th e ir p h y s ic o - c h e m ic a l c h a ra c te ris tic s.
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Physical c h a r a c te r is tic s o f the feeds:
T h e p h y sical c h a r a c te r i s tic s o f th e th r e e feed s p r e p a re d b y in c o rp o ra tin g three levels o f dried c a b b a g e w a s t e at 5, 10 a n d 15% a n d d e s ig n a te d as C W I, C W II and C W III are e l a b o r a te d in T a b l e 7. A s trik in g ch ara cte ristic w a s the u n ifo rm ity in p e lle t s h a p e m a in ta in e d b y th e three feed s, w h i c h m a y be attrib u ted to th e fm e te x tu r e o f d r ie d p o w d e r e d c a b b a g e w a s te . T h e fe e d w^ith 5 % in c o rp o ra tio n w'as p a le b r o w n in c o lo r w h ile the feed s in c o r p o ra te d w ith 10 and 15% c a b b a g e w a s t e h a d a d a r k e r b r o w n colo r w ith a g re e n ish tin g e w h ic h in c r e a s e d w ith le v e l o f in c o r p o r a tio n o f c a b b a g e w a s te . T h e control feed w as brow^n in c o lo r a n d h a d a s lig h tly u n e v e n te x tu re . It e x u d e d a s tro n g fishy o d o u r w h i l e th e feed s p r e p a r e d f r o m c a b b a g e w a s te e x u d e d a m ild odour.
Table 7. Physical characteristics of the control and experimental feeds compounded utilizing dried unfermented cabbage waste
FEED P E L L E T SIZE PHY SICAL APPEA RA NCE
Control 2.Ox 3.0 mm Dark brown uneven texture
CWI 2.0x 3.0 m m pale brow n fme and compact
CWII 2 .0 x 3 .0 mm dark brown greenish tinge fme
and compact CWIII
...
2 .Ox 3.0 mm dark brown greenish tinge fme and compact
H y d r o s t a b i l i t y o f t h e d r i e d u n f e r m e n t e d C a b b a g e W a s t e f e e d s:
T he h y d r o s ta b ilty o f th e th re e fe e d s p r e p a r e d in c o rp o ra tin g 5,10 a n d 15%> d rie d cab b ag e w 'aste a n d a ls o o f th e c o n tro l f e e d s is giv en in fig u re 1. T h e th re e feeds viz. C W I, C W I I a n d C W I I I p r e p a r e d in c o rp o ra tin g 5, 10 a n d 15% o f c a b b a g e w aste r e s p e c tiv e ly w e r e q u ite s ta b le in s e a w a te r w ith 7 8 - 8 0 % dry m a tte r
rem ain in g at th e e n d o f o n e h o u r. T h is d id n o t d ec re a s e a n y further, so th a t b y the end o f 4 h o u rs o n l y a m a r g in a l a d d itio n a l loss r a n g in g fro m 5 to 8 % w a s re c o rd e d in the c a s e o f th e s e feed s s h o w i n g th e m to b e h ig h ly stable d e s ig n e d sp ecifically for s h r i m p f e e d in g . T h e c o n tr o l fe e d g av e sim ila r w a te r stability.
P ro x im a te c o m p o s itio n o f the u n f e r m e n t e d feeds:
F e e d s C W I, C W I I a n d C W I I I r e c o r d e d m o i s t u r e c o n te n ts r a n g in g b e tw e e n 3.3 to 4 .2 % ( T a b le .8). F e e d C W I p r e p a r e d in c o r p o r a tin g 5 % c a b b a g e w aste r e c o r d e d a p ro te in c o n te n t o f 4 1 . 0 1 % as c o m p a r e d to 4 3 .3 9 % o f th e control feed, w h ile fe e d s C W I I a n d C W I I I p r e p a r e d u s in g 10 a n d 15% o f th e dried c a b b a g e w a s te r e c o r d e d v a lu e s o f 3 8 . 6 0 % a n d 3 7 . 0 3 % re s p e c tiv e ly as th e cru d e p ro te in c o n te n ts . T h e f e e d s p r e p a r e d fro m u n f e r m e n te d c a b b a g e w a s te h a d lo w e r fat c o n t e n t r a n g i n g b e t w e e n 5 .1 5 % fo r fe e d C W I I I to 5.87%> for feed C W I, th a n th e c o n tro l fe e d w h i c h r e c o r d e d a fat c o n t e n t o f 6 .1 8 % . H o w e v e r , the r e v e rs e w a s the c a s e fo r c r u d e f ib e r c o n te n ts w h e r e in , h ig h e r v a lu e s o f 2 .64 % , 3 .1 0 % an d 3 .1 0 % r e s p e c t iv e l y w e r e o b ta in e d fo r f e e d s , C W II a n d C W III as c o m p a r e d to 2 .0 5 % c r u d e fib e r c o n t e n t o f th e c o n tro l feed.
Table«^: P r o x i m a t e chem ical com position of the control an d e x p e rim e n ta l feeds c o m p o u n d e d utilizing v a ry in g c o n c e n tra tio n s of d rie d cabbage waste.
N U T R IE N T
F E E D S ’’^
Control CWI CWII CWIII
Dry matter 97.048 96.597 95.774 96.704
Moisture 2.95 3.403 4.226 3.296
Crude Protein 43.391 41.011 38.903 37.033
Ether extract 6.182 5.871 5.687 5.152
Crude fiber 2.051 2.637 3.104 3.097
Ash 10.771 10.382 9.958 9.103
Acid insoluble ash (A IA ) 0.616 0.508 0.463 0.216
NFE** 34.653 36.696 38.421 42.319
•V alu es expressed on dry matter basis and average o f three estimations carried out in triplicate
** N itrogen Free Extractives- calculated as (100-%crude protein +crude fat+ ash + crude fiber+ m oisture)
A s h c o n t e n t d e c r e a s e d s lig h tly in th e c a b b a g e w a s te in c o rp o ra ted feeds w ith fe e d s C W I , C W I I a n d C W I I I r e c o r d i n g 10.38% , 9 .9 6 % a n d 9 .1 0 % ash. T h e ash c o n t e n t o f t h e c o n tro l fe e d w a s 1 0 .7 7 % . A m a r g in a l in crease w a s also o b ta in e d in th e c a r b o h y d r a te c o n te n ts o f th e feed s p r e p a r e d in c o rp o ra tin g 5 to 15% c a b b a g e w a s t e . H e r e c a r b o h y d r a te c o n te n t in c re a s e d fro m 3 4 .6 5 % in the control fe e d to 3 6 . 7 0 % u p o n 5 % in c o r p o r a tio n o f c a b b a g e w aste. T h is in c re a s e d further f ro m 3 8 . 4 2 1 % u p o n in c o r p o r a tio n o f 10% c a b b a g e w aste to a h ig h e r value o f 4 2 . 3 2 % u p o n in c o rp o ra tio n o f 15% c a b b a g e w a s te . A c id in s o lu b le ash con ten t d e c r e a s e d s ig n ific a n tly fro m 0 .6 2 % o f th e c o n tro l feed to a v a lu e o f 0 .2 2 % in fe e d C W I I I in c o r p o ra te d w ith 1 5 % c a b b a g e w a s te .
S o / i d Sfflfp f p r m g n t a t i o n o f C a b b a g e w a s t e P e t r i p l a t e a n d in C o n i c a l f l a s k
Plate V: Solid State Fermentation o f cabbage waste in petri plate and in conical flask w ith B. coagulans.
feto p fa(tao ,U iij:« iiical l l a s k ^
^ale VII: Solid State Fermentation o f Gingelly oilcake in petri plate and in conical
Jask with D. coagulans.
O n e n o n - c o n v e n t i o n a l p la n t in g r e d ie n t viz. c a b b a g e w a s te an d t w o an d co nv entio n al p la n t in g r e d ie n t s viz. C o tt o n s e e d c a k e a n d g in g e lly w ere s u b je c te d separately to S o lid S ta te f e r m e n ta tio n u s i n g b a c te r ia {B.coagidans) a n d F u n g i {Beauveria sp.). In th e c a s e o f f e r m e n ta t io n o f v a r io u s su b stra te s c a r r ie d o u t em p lo y in g b a c te r ia , t h e r e w a s n o p h y s i c a l c h a n g e in a p p e a r a n c e ap a rt f r o m th e ferm ented o d o u r o b t a i n e d u p o n c o m p le t e f e r m e n ta tio n a f te r 48 hrs (P la te V to VII). D r y m a tte r c o n t e n t o f d rie d f e r m e n t e d c a b b a g e w a s t e in c re a se d to 9 9 .6 7 % while th e re w a s a c o n c o m i t a n t d e c r e a s e in m o is tu r e to 0 .3 3 % ^ ( P la te V III, T a b le 9 )^A m ild in c r e a s e w a s o b ta in e d in th e c r u d e p ro te in w ith a v alu e o f 1 6 .4 5 % being r e c o r d e d a f t e r S S F w ith B .coagulans. F a t c o n te n t o f fe rm e n te d C a b b a g e w aste s h o w e d n o c h a n g e w ith a v a lu e o f 1 .5 5 % b e in g o b ta in e d . A slig h t d e c r e a s e w'as o b s e r v e d in th e c a s e o f c r u d e fib e r an d a sh c o n te n ts o f B.coagulans fe rm e n te d c a b b a g e w a s te , w h ic h r e p o r te d v a lu e s o f 1 1 .7 4 % for th e f o r m e r and 13.59% fo r th e la ter. A v a lu e o f 5 6 .3 4 % w as re c o rd e d as the c a r b o h y d r a te content w^hile a c i d in s o lu b le a sh r e m a i n e d static at 0 .0 9 % . In the ca s e o f d rie d ferm e n ted c o t t o n s e e d c a k e (P la te I X ) w ith 9 9 .2 8 % d ry m a tte r a n d 0 .7 2 % m oisture, c r u d e p r o t e i n c o n te n t r e c o r d e d a m a rg in a l in c r e a s e re c o r d in g a v a l u e o f 29.68% . T h e r e w a s a c o n c o m it a n t d e c r e a s e in lip id w h e r e 4 .4 9 % w as r e c o r d e d as the c ru d e fat c o n t e n t ( T a b le 9).
d r i e d FF.RMF?MTirn
Plate IX ; D ried C o lto n seed cake after being subjected to fermentation with
B. c o ^ itk m sNutrient SU B S T R A T E *
C W CSC GOC
Dry matter 99.668 99.276 99.599
Moisture 0.332 0.724 0.401
Crude Protein 16.453 29.68 39.589
Ether extract 1.548 4.49 8.54
Crude fiber 11.739 17.28 2.93
Ash 13.588 7.707 15.421
Acid insoluble ash (A IA ) 0.088 0.237 1.299
NEE** 56.34 40.519 33.451
* Values expressed on dry m atter basis and average o f three estimations carried out in triplicate.
** Nitrogen Free Extractives- calculated as (100-%crude protein +crude fat+ ash + crude fiber+m oisture)
r e r m e n l a t i o n o f c o tto n s e e d c a k e w ith B .coagulans resu lted in a m ild increase in b o th c r u d e f ib e r ( 1 7 .2 8 % ) a n d ash c o n te n ts (7 .7 1 % ). T h e re w a s no sig n ific a n t c h a n g e in th e c a r b o h y d r a te a n d A I A c o n te n ts o f cotton s e e d cak e fe rm e n te d w ith B .coagulans as a v a l u e o f 4 0 .5 2 % w a s r e c o r d e d for th e fo rm e r a n d a v a lu e o f 0 . 2 4 % f o r th e later.
S o lid S ta te F e r m e n ta t io n o f g in g e lly o ilc a k e re c o rd e d a d ry m a tte r co n ten t o f 9 9 . 6 % a n d m o is tu r e c o n t e n t o f 0.4 % . P r o te in in c re a se d in this c o n v en tio n al in g r e d ie n t fro m th e u n f e r m e n te d c o n te n t o f 34 .4% to j 9 . 5 9 % (T ab le 9, P la te X )). T h o u g h c r u d e fat. C ru d e fib e r a n d ash re p o r te d m ild increases r e g a r d in g v a l u e s o f 8 .5 4 % , 2 . 9 % a n d 1 5 .4 2 % r e s p e c tiv e ly a d e c r e a s e w a s obser\'ed in th e c a s e o f c a r b o h y d r a t e c o n te n t w ith a v a lu e o f 33.45%> b e in g r e c o rd e d as th e c a r b o h y d r a te o t g in g e lly o ilc a k e a tte r ferm e n ta tio n w ith B.coagulans. T h e a c id in s o lu b le a s h c o n t e n t h o w e v e r r e m a in e d unaltered .
28
