A q uacultural Engineering 3 ( t 984) 289-301

The Utilization of Clam Waste Meal as a Protein Source

for Rainbow Trout Salmo gairdneri

*C.L. Goodrich, *S.M. Barnett, +G. Levine and *K.L. Simpson

* Department of Food Science and Technology', Nutrition and Dietetics,
University of Rhode Island, Kingston, Rhode Island 02881. USA
Blount Seafood Corp., Warren, Rhode Island 02885. USA

A BS TRA CT

SolM waste recovered from clam processing wash water was evahtated lbr
its utilizatkm as a replacement protebz for fish meal. Three diets, diets 1
and 2 fi)rmulated to simulate the Oregon Moist Pellet {OMP) and diet 3
{Rangen), were fed to rabzbow trout (Salmo gairdneri)for eight weeks. The
effects o f the clam waste meal on fish growth, food conversion, carcass
composition attd whole body fatO' acM content were determhwd. The
clam waste diet supported mcLt'imttm growth attd food conversiott. Carcass
attalysis o f moisture, proteht and lipid contents were similar among the
moist feeds. The d#tary levels o f 20 : 5w3 + 22 : 6w3 and total poO'-
unsaturated fatty acids (PUFAs} were significantly higher bz the clam
waste diet pellet.

INTRODUCTION

Traditionally, fish meal has been the dominant protein source in fish
leeds. Fish meal is becoming increasingly expensive and in short supply;
therefore, a strong economic incentive exists to find alternative protein
sources (Fowler and Banks, 1976; Foltz et al., 1982; Jackson er al.,
1982).
Plant proteins are commercially available and are less expensive as
fish meal supplements. However, many factors affect the nutritional
quality o f plant proteins, and problems such as inferior amino acid
composition, antinutritional and toxic factors and processing conditions
must be overcome when using these ingredients in diet formulations
289
Aquacultural Engineering 0144-8609/84/$03.00 © Elsevier Applied Science
Publishers Ltd, England, 1984. Printed in Great Britain
290 C. L. Goodrich, S. M. Barnett, G. Levine, K. L. Simpson

(Rumsey, 1973). Other protein feedstuffs of animal origin have suc-

cessfully replaced fish meal. Disappointingly, most of these sources are
scarce and/or as expensive as fish meal (Viola et al.. 1981, 1982).
The dam processing industry generates an appreciable amount of
waste. Wash water in clam processing plants contains small particles of
clam tissue and soluble solids. Elimination of this organic waste not
only magnifies disposal problems, but also results in the non-utilization
of a valuable protein resource (Joh and Hood, 1979). There is a need to
develop new treatment methods and productive means for the utiliza-
tion of clam wastes. Clam waste has been shown to have a relatively
high protein content necessary for its inclusion in fish feeds (67.9% on
a dry weight basis; Hang e t a l . , 1980).
The present study was conducted to evaluate clam waste meal as a
fish meal replacement in salmonid diets. The effects of the clam waste
on fish growth, food conversion, carcass composition and whole body
fatty acid content were determined.

MATERIALS AND METHODS

Three diets, two simulating the OMP, a commercial moist fish feed
(Table 1), and Rangen, a commercial dry feed, were used in the present
study. The moist diets differed only in the replacement of fish meal
with clam waste meal.

Source of clam waste meal

Solid waste was obtained from a commercial clam processor located

in Warren, Rhode Island. The material was collected by the sedimenta-
tion of clam processing wash water. Recovered waste was freeze-dried,
ground and stored at --20°C under nitrogen. Duplicate samples of the
clam waste meal were subjected to proximate analysis, the results of
which are shown in Table 2. Protein, crude fat, moisture, ash and crude
fiber were determined by standard methods (AOAC, 1975). Proteins
were acid hydrolyzed in 6-0 N HCI by the procedure of Spackman et al.
( 1958) as modified by Niederwieser and Pataki ( 1971). The amino acid
composition (Table 2) was measured by ion-exchange chromatography
using an Nc-2P Technicon auto-analyzer (cf. Seidel et al., 1980).
 Utilization o f clam waste meal as protein source for rainbow trout 291

TABLE 1
Composition of OMP Diets

hzgredien t Perce, tage o f total diet

Diet 1 Diet 2

Fish meal (Menhaden) 29.0 -

Clam waste meal - 29-0
Cottonseed meal 15.0 15,0
Wheat germ meal 8.9 8-9
Dried whey 5-0 5.0
Corn distillers dried solubles 4.0 4.0
Wet fish component a 30-0 30.0
Fish oil (Menhaden) 3.0 3.0
Soybean oil 3-0 3.0
Choline chloride (70% liquid) 0.5 0.5
OMP vitamin premix b 1.5 1.5
OMP mineral premix c O-I O. I

a Ground, pasteurized flounder.

b Contains in mg kg-~ premix: L-ascorbic, 59400; D-biotin,
40: viu, min BI 2.4.0: alpha-tocopherol acetate. 33 440 [U"
folic acid, 847; myoinositol, 17 600; menadione sodium
bisulfite. 1199; niacin. 12540: D-calcium pantothenate,
7652: pyridoxine HCI: 1430: riboflavin, 3520: thiamine
mononitrate, 1712.
c Contains in g kg-1 premix: ZnSO4, 185: MnSO,;, 207:
FeSO,:.7H20, 50: CuSO4, 4.0: KIO3, 0 4 : CoSO4.7H~O, 25.

Diet formulation

T h e OMP diets (I and 2) were m i x e d and pelleted in a H o b a r t c o m -

mercial m e a t g r i n d e r using a 3 / 3 2 in die. T h e pellets were blast frozen
and stored at - - 2 0 ° C until fed. R a n g e n (diet 3) was p u r c h a s e d f r o m
Zeigler B r o t h e r s Inc. ( G a r d n e r s , P e n n s y l v a n i a ) and used as supplied.
292 C. L. Goodrich, S. M. Barnett, G. Levine, K. L. Simpson

TABLE 2(a)
Proximate Composition of Clam Waste Meal

Component Percott

Crude protein (N x 6.__) 70.0

Crude fat 9.8
Ash 11 -1
Moisture 3.0
Crude fiber 0-3
NFE a 5-8

a Nitrogen-free extractive determined by

difference.

TABLE 2(b)
Amino Acid Content of Clam Waste Meal

Amino acM g 100 g-1 sample

Alanine 3.6
Arginine 6. I
Aspartic acid 5.9
½ Cystine 0.4
Glutamic acid 7.8
Glycine 1-4
Histidine 1.5
lsoleucine 3-0
Leucine 4.5
Lysine 6.1
Methionine 1-6
Phenylalanine 2.5
Proline 3.2
Serine 3.7
Threonine 3.6
Tryptophan a
Tyrosine 2.6
Valine 3-4

a Not determined.
 Utilization o f clam waste meal as protein source/'or rainbow trout 293

Diet analysis

The proximate composition of the fish diets was determined by AOAC

(11975) methods. The total caloric contents were calculated using
energy values of 3.9, 8-0 and 1-6 kcal g-~ for protein, fat and carbo-
hydrate, respectively (National Academy of Sciences, 1973). Dietary
amino acids were analyzed as described above.
Lipids were extracted by the method of Btigh and Dyer (1959) as
modified by Kates (1972) for fatty acid analysis, blethyl esters were
prepared by saponification with 0-5N potassium hydroxide and
methylation with 14% boron trifluoride-methanol (Morrison and
Smith, 1964). Fatty acid methyl esters (FAblE) were analyzed on a
single column Varian Aerograph 1700 gas-liquid chromatography unit
operated at 200°C isothermally and equipped with a flame ionization
detector. The FAMEs were separated on a 10% SP-2330 column sup-
plied with 30 ml rain -~ nitrogen as the carrier gas. A 37- ethylene glycol
succinate polyester-Z (EGSP-Z) column was used to confirm fatty acids
of similar retention time on the SP-2330 column. FAMEs were identi-
fied with an electronic integrator (Hewlett Packard 3380 A) supplied
with the relative retention times of methyl ester standards (cf. Schauer
er al., 1980), Cod liver oil was used as a secondary reference standard
(Ackman and Burgher, 1965).

Feeding study

Rainbow trout S a l m o g a i r d n e r i were purchased from the Plymouth

Rock Trout Co. (Plymouth, Massachusetts). Following an acclimation
period of four weeks, fish averaging 70.9 g were counted into groups of
25 and randomly distributed into each of six tanks. Rearing units were
45-gallon circular fiberglass tanks supplied with an average water flow
of 4 liters min-L Oxygen content, temperature and water pH were
monitored biweekly. The mean dissolved oxygen level was 7.4 ppm,
temperature ranged between 12 and 15°C and the mean pH was 5.9.
Each diet was fed to fish in duplicate tanks for the duration of eight
weeks. Fish were fed twice daily, six days a week, with food offered as
long as the fish continued to actively feed. The fish were weighed every
two weeks and average weight data were subjected to analysis of vari-
ance to determine if differences in means were statistically significant
294 C. L. Goodrich, S. M. Barnett. G. Levine. K. L. Simpson

at the 5% level. The tables used for statistical analysis were from
Snedecor and Cochran (1967). F o o d consumption and mortality were
recorded daily.

Carcass analysis

At the termination of the study, five fish were randomly selected from
each group for proximate analysis and whole body lipid analysis. Fish
samples from each tank were pooled and homogenized in a Waring
blender. Moisture and ash contents were determined immediately: the
remainder o f the sample was lyophilized and stored under nitrogen for
crude lipid and protein analysis. Whole body lipids of the wet tissue
homogenates were extracted and fatty acids analyzed by gas-liquid
chromatography as described above.

RESULTS AND DISCUSSION

The proximate composition of the fish diets is given in Table 3. The

protein content was 38% with the exception of diet 1 which provided
33% protein. Crude fat contents ranged from 7.9% in diet 3 to 13.25';
in diet 1. The total caloric value was similar among the three feeds.

TABLE 3
Proximate Composition of Diets (All Values on As-fed Basis)

Diet Moisture Chide Crude Ash Crude NFE b Total

number (%) prote#z a fat (%) fiber cah~ric
(%) (%) (%) vahte
( kcal 1 O0
g-l diet)

l 28.8 32-6 13.2 8.1 1-7 15.6 257,7

2 24.8 37-5 11.9 6-1 1.7 18-0 270.3
3 10.2 37.7 7-9 8-8 2-2 33.2 263.3

a N x 6-25.
b Nitrogen free extractives determined by difference.
 Utilization o f clam waste meal as protein source ]br rainbow trout 295

TABLE 4
Amino Acid Composition ot" Diets [g 100 g-~ Diet, As-fed Basis)

Diet I Diet 2 Diet 3 Requirement a

Amino acid
Alanine 2.0 1.6 2.3
Arginine 2.9 2.4 2.1 2.4
Aspartic acid 2.5 2.3 3.0
Cystine 0-I 0.1 -
Glutamic acid 4.4 4.2 4-6
Glycine 2.1 1.6 2.2
Histidine 0.9 0.7 1-0 0.7
Isole ucine 1.3 1-0 0.8 0.9
Let, cine 2.1 2-3 2.7 1.6
Lysine 2-0 2-1 2.0 2.0
Methionine ~ 0-7 0.7 0.6 0.5
Phenylalanine c 1-4 1.2 1.6 2.1
Proline 2.0 1.9 1.8
Serine 1.3 1.6 2.1
Threouine 1.3 1.4 1.2 0.9
Tryptophan a - - - 0.2
Tyrosine 0.9 1.0 0.9
Valine 1-6 1.2 1.4 1.3

a Values for Chinook salmon fingerlings receiving 40% protein (Nat. Acad. Sci.,
1973).
t, Ii1 the presence of 1% cystine.
c In the absence of tyrosine.
a Not determined.

No a p p a r e n t d i f f e r e n c e s were s h o w n in the essential a m i n o acid

c o m p o s i t i o n o f the diets (Table 4). T h e essential amino acid require-
m e n t s for C h i n o o k salmon were m e t (Nat. Acad. Sci., 1973) w i t h the
e x c e p t i o n o f arginine and isoleucine in diet 3 and valine in diet 2. Pre-
cautions were n o t t a k e n t o p r e v e n t cystine f r o m o x i d a t i o n in acid
h y d r o l y s a t e s ; t h e r e f o r e , with partial d e c o m p o s i t i o n it can be supposed
that the c o n t e n t o f this a m i n o acid was originally higher (Moore, 1963).
All o f the diets were d e f i c i e n t in p h e n y l a l a n i n e ; h o w e v e r , t y r o s i n e has
a sparing a c t i o n o n p h e n y l a l a n i n e (Satia and B r a n n o n , 1975).
296 C. L. Goodrich, S. :~I. Barnett, G. Levine, K. L. Simpson

~ 12C

BC

i I I
0 2 4 6 8
Time (weeks)

Fig. 1. Growth curves of rainbow trout fed diets for eight weeks. Curves (from
top to bottom) represent fish fed diets 2, 1. and 3 respectively.

No mortalities were observed throughout the eight week feeding

study. A growth-response curve for each diet (Fig. 1) was obtained by
plotting the mean weight of replicate fish treatments measured initially
and biweekly thereafter. Differences in growth rates among the diets
became apparent after the fourth week. The best overall growth per-
formance was observed in fish fed the clam waste meal diet. Analysis
of fish weights showed statistical differences between diet treatments
at the 5% level. Fish fed diets 1 and 2 had significantly higher weight
gains than fish fed diet 3. Weight gain data between the two OMP
fommlations did not show statistical differences.
Food conversions, on a dry weight basis, of fish f e d diets I and 2,
were significantly better than those fed diet 3 (Table 5). Good food
conversion values and tow incidence of several diseases have been
observed in fish culture on the OMP (Ghittino, 1972).
 Utilization or" clam waste meal as protein source/or rainbow trout 297

TABLE 5
Average Body Weights, Percent Gain, Food Conversion. and Feeding Rates for
Rainbow Trout Fed Diets For Eight Weeks

Diet Average body weight (g) Percent Food Feediw rate b

mlmber gain conversion a {% body
Inisial Final weight)

1 70.8 140.1 97.9 1-18 1.9

"~ 71.5 151.5 111.9 1.01 1-9
3 70.4 126.0 79.0 1.38 i.9

a Food conversion = food, dry weight (g)/weight gain (g).

b Calculated on a dry weight basis.
Weight and food conversion are the mean values for replicate tanks.

TABLE 6
Moisture, Protein, Lipid, and Ash Contents of the Carcasses of Rainbow Trout
Fed for Eight Weeks

Diet Jmmber Moisture % in whole bodies offish

Protein Lipid .4 sh

I 685 16-8 11.2 2.3

2 68-5 16.6 11-3 2-5
3 69.6 17.5 9-9 2-5
Initial 71.2 17-0 7.9 2-4

Carcass analysis o f m o i s t u r e , p r o t e i n and lipid c o n t e n t s at the termi-

nation o f the feeding s t u d y were the same for diets 1 and 2 as s h o w n in
Table 6. Fish fed diet 3 had a slightly higher carcass p r o t e i n c o n t e n t .
T h e m o i s t u r e and c r u d e lipid levels were inversely related, with mois-
ture c o n t e n t increasing and lipid c o n t e n t decreasing, as previously
d o c u m e n t e d ( J a u n c e y , 1982). Ash c o n t e n t in the fish carcasses did not
vary b e t w e e n the diet t r e a t m e n t s .
T h e fatty acid c o m p o s i t i o n o f fisil b o d y lipids reflected the f a t t y
acid c o m p o s i t i o n o f the diets as s h o w n in Table 7. T h e r e was a positive
298 C. L. Goodrich, S. M. Barnett, G. Levine. K. L. Simpson

TABLE 7
Fatty Acid Composition of Dietary l_ipids and Total Body Lipidsa ~Percent)

Fatta, acid Diet 1 Diet 2 Diet 3

Diec Fish Diet Fish Diet Fish

12 0 0.06 0-19 - 0.20 2.04 0.90

14 0 5-85 3-51 3.8l 2.75 4.40 3.35
15 "0 0.42 0.30 0.28 0.27 0.44 0.34
16 0 19.53 16-86 18.66 18.15 22-23 18.23
16" lw7 7.51 7.28 7.28 7-13 5.19 8.54
16 " 2w4 0-60 - 0.31 - 0.33 -
18 0 3.10 4.99 3.61 5-78 5-59 5-40
18' lw9 19.49 26.12 16-53 25-18 20-08 33.09
18 " 2w6 ._.78 19.o9 25.57 18.,.~
"~" _ _
"~6"-~8 13-43
18 awa
" " 5.10 - 4.64 - 3.71 -
18 4w3 ,.15 1.18 2.52 1.10 0.82 0.80
20 I w9 - 4.59 - 4.49 - 3.42
20 2w6 _ 0-.9 a"~ 0-43 1.05 - 0.88
20 3w6 - 0.29 - 0.33 0.08 0.43
20 4w6 0.54 0.67 0.29 0-54 0.12 0.27
20 3-W . ~" 6-71 3.02 9.90 3.29 3.55 1.52
22 1 1.63 I .79 0-83 1.45 1-24 1.74
22 5w3 0.38 0-75 0.60 1.04 0.15 0.32
22 6w3 3.87 7.86 4.50 8-65 3.76 7.13
24"1 0.34 0.38 0.43 0-48 0-07 0-40
Total saturated
fatty acids 28.96 25.85 ..6-40"
"~ 27.15 34.70 28.22

a Average analysis of duplicate tanks.

c o r r e l a t i o n b e t w e e n d i e t a r y levels o f 14 • 0, 15 • O, 16 • 0, 18 " I, 18 • ~
18" 4, 2 0 4, 20 • 5, 22 • 1, 22 • 5, 22 " 6 a n d 2 4 • 1 a n d fish lipid levels
f o r all e x p e r i m e n t a l g r o u p s .
The total saturated fatty acid ( S F A ) content in fish b o d y tipids
r e m a i n e d fairly c o n s t a n t at 27% r e g a r d l e s s o f t h e high S F A level in d i e t
3 ( 3 4 - 7 % ) . T h i s f i n d i n g is c o n s i s t e n t w i t h t h a t o f Y u e t al. ( 1 9 7 7 ) . T h e y
r e p o r t t h a t p a r t i a l r e p l a c e m e n t o f h e r r i n g oil b y a n i m a l l a r d , w h i c h
c o n t a i n s a high p e r c e n t a g e of SFAs, does not increase the s a t u r a t i o n of
 Utilization of" elalq'z waste meal as protein source/br rainbow trout 299

TABLE 8
Essential Fatty Acid Contents (%1 of Diets

Diet number Total w3 PUFA 20 : 5 w 3 + 22 . 6w3 18 . 3w3

(~) ,-"

I 2-40 1.40 0-67

2 2.64 1.71 0.55
3 0-95 0.58 0.29

fish lipids. Apparently, a mechanism may exist in fish to regulate and

maintain a proper level of body lipid saturation.
The total w3 PUFAs, 2 0 5 w 3 + 2 2 " 6 w 3 , and 1 8 3 w 3 contents
were calculated for the diets, tile results of which are given in Table 8.
Diet _~ is higher in total w3 PUFAs and 2 0 ,gw3 + .;_~" 6w3 contents
than the other diet groups. Linolenic acid ( 1 8 : 3 w 3 ) levels are some-
what low in all three of the diets, However, Takeuchi and Watanabe
(1977) showed that the replacement of 0 . 5 ' 5 linolenate with either
0.55~ eicosapentaenoate ( 2 0 : 5 w 3 ) o r 0.57, docosahexaenoate (22: 6w3)
markedly increases the growth rate of rainbow trout. They demon-
strated that 2 0 5 w 3 in addition to 2 " 6 w 3 has an EFA efficiency
higher than 1 8 " 3 w 3 . The best weight gain was observed in fish fed
diets containing 0.25% each o f 20 : 5w3 + 22 : 6w3, respectively, indi-
cating an additive effect between these fatty acids on tile growth
response of rainbow trout. It is postulated that the superior growth
response shown in diet 2 was partially due to the higher 2 0 : 5 w 3 +
2 2 : 6 w 3 composition.

CONCLUSIONS

This study shows that clam waste meal can be utilized by rainbow trout
as a protein source. Tile production of clam waste meal to be used as
a feed ingredient illustrates a potential solution to tile problem of waste
management for the clam industry. Waste material which is unfit for
human consumption can be recycled through fish, to be sold as an
acceptable human food.
300 C. L. Goodrich, S. M. Barnett, G. Levine, K. L. Simpson

ACKNOWLEDGEMENT

Part of this work was presented at the 43rd Annual Meeting o f the
Institute o f F o o d Technologists, New Orleans, Louisiana, 19-22 June
1983. This study was s u p p o r t e d by Sea G r a n t contract no. N A 8 1 A A -
D-00073.

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