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SID 5
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Research Project Final Report
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SID 5 (2/05)
Project identification
1.
Defra Project code
2.
Project title
FC0931
Laboratory assessment of samples involved in
substitution of fish meal with vegetable proteins in cod
diets
3.
Contractor
organisation(s)
SAMS Ardtoe, Ardtoe Marine
Laboratory, Acharacle, Argyll
PH36 4LD
54. Total Defra project costs
5. Project:
Page 1 of 16
£
7075.00
start date ................
16 October 2004
end date .................
15 February 2005
6. It is Defra’s intention to publish this form.
Please confirm your agreement to do so. ................................................................................... YES
NO
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Executive Summary
7.
The executive summary must not exceed 2 sides in total of A4 and should be understandable to the
intelligent non-scientist. It should cover the main objectives, methods and findings of the research, together
with any other significant events and options for new work.
Authors and partners:
J. Treasurer, C. Cutts
SAMSardtoe, Ardtoe Marine Laboratory, Ardtoe, Acharacle, Argyll, PH36 4LD
I. Pike, Technical Director, IFFO, 2 College Yard, Lower Dagnall Street, St Albans, Herts AL3
4PA, UK
R. Alderson, BIOMAR, North Shore Road, Grangemouth Docks, Grangemouth FK3 8UL
T. Galloway, BIOMAR, Norway
N. MacDonald, EWOS, Westfield, Bathgate, West Lothian EH48 3BP
Jon Arnason, Laxa, Laxa Feedmill Ltd, Krossanes, IS-603 Akur, Iceland
S. Albrektsen, SSF, Fiskeriforskning, Norsk institutt for fiskeri- og havbruksforskning AS,
Kjerreidviken 16, N-5141 Fyllingsdalen, Norway
G. Bell, V. Karalazos, Institute of Aquaculture, University of Stirling FK9 4LA
SID 5 (2/05)
Page 2 of 16
1. This study expands on report Defra FC0930 detailing the effects of substitution of fish meal
with vegetable protein on growth of cod. The results of analyses of muscle, liver, bone
samples, essential fatty acids and pathological effects of substitution are examined. The
methods used for faeces extrusion and mineral analysis were developed through the study
and may serve as a standard for future studies.
2. Apparent digestibility coefficients (ADCs) for fat, protein and starch decreased with inclusion
of fullfat Soya but increasing levels of substitution did not appear to affect the ADC further.
3. Regarding bone mineralisation, only zinc content was affected by dietary treatment,
increasing significantly with increasing levels of fullfat Soya.
4. The inclusion of fullfat Soya did not have a marked effect on the whole body composition.
Likewise, the whole body nutrition did not appear to be affected by the levels of fullfat Soya
tested in the trial. The inclusion of fullfat Soya in the diet reduced both protein and energy
efficiencies, but did not appear to be affected by the level of fullfat Soya.
5. The fatty acid profile of the liver and the diet were almost identical. All dietary treatments
produced high EPA and DHA levels in the livers, although linoleic acid levels were high. The
levels of the latter would preclude livers being used for cod liver oil. The fatty acid
composition of muscle changed as the EPA and DHA content of the diet fell. The differences
were not significant. As muscle cells are normally of consistent composition in terms of total
lipid content and fatty acid makeup, this unexpected finding requires further investigation
(Bob Ackerman, pers. comm., 2005).
6. Fish fed fullfat Soya at 36% substitution showed only mild posterior gut inflammation and the
severity was significantly less than in Atlantic salmon fed similarily substituted diets.
However, these effects may affect performance following diet substitution and the effects of
fullfat Soya on the immune system need to be examined further.
7. In conclusion, this study demonstrates that cod can utilise substituted raw materials in the
diet but there is a slight effect on cod performance. A cost benefit analysis should be
undertaken of using whole Soya as a protein and oil source.
Project Report to Defra
8.
As a guide this report should be no longer than 20 sides of A4. This report is to provide Defra with
details of the outputs of the research project for internal purposes; to meet the terms of the contract; and
to allow Defra to publish details of the outputs to meet Environmental Information Regulation or
Freedom of Information obligations. This short report to Defra does not preclude contractors from also
seeking to publish a full, formal scientific report/paper in an appropriate scientific or other
journal/publication. Indeed, Defra actively encourages such publications as part of the contract terms.
The report to Defra should include:
 the scientific objectives as set out in the contract;
 the extent to which the objectives set out in the contract have been met;
 details of methods used and the results obtained, including statistical analysis (if appropriate);
 a discussion of the results and their reliability;
 the main implications of the findings;
 possible future work; and
 any action resulting from the research (e.g. IP, Knowledge Transfer).
SID 5 (2/05)
Page 3 of 16
2. Introduction
As cod, a carnivorous species, has a high protein requirement, farmed cod are fed a diet with around
60% fish meal. Replacing part of this fish meal with vegetable protein would reduce the cost of the
feed and the amount of wild fish used indirectly in the feed. However, vegetable proteins contain
carbohydrates which are poorly utilised and they also contain anti-nutritional factors. Most of these
have been removed by improvements in processing. Carbohydrates can also be removed but the cost
is expensive.
The present work is to analyse fish muscle, liver, and bone samples already collected from a previous
Defra study FC0930 to determine changes in proximate composition and essential fatty acids. Any
pathological changes associated with dietary substitution were also examined.
The study was conducted by SAMS Ardtoe in conjunction with the International Fish Meal and Fish Oil
Association (IFFO), three fish feed companies EWOS, BioMar, LAXA, and SFF* Laboratory in Norway
(* now the Norwegian Institute of Fisheries and Aquaculture Research). Laboratory analyses were
performed by the Institute of Aquaculture, Stirling and histology by the Fish Vet Group, Inverness.
The relevance of the study is to ensure sustainability of feeds for marine finfish species given the
rapidly expanding nature of the fish farming sector. This expansion may be extended from primarily
salmon farming in north temperate areas to alternative marine finfish species and this area of growth
may be encouraged in many respects by the decline in marine finfish species. This may place further
pressure on the cost effective supply of fish meal.
3. Methods
The methods used in the full study were reported in DEFRA report FC0930, project ”Substitution of fish
meal with vegetable protein on cod diets.” The methods used for faeces extrusion and mineral
analysis were developed through the study and may serve as a standard for future studies.
Proximate analysis; Proximate analysis of feed and tissues was conducted using standard methods
described by the AOAC (1995).
Moisture; Moisture was determined by heating the weighed sample at 110oC for 16h or until constant
weight was achieved.
Protein; Crude protein (N x 6.25) was determined by the Kjeldhal method after acid digestion.
Oil; Crude fat was determined in diets by acid hydrolysis using a Soxtec System 1047 hydrolysing unit
(Tecator Application note 92/87 followed by exhaustive Soxhlet extraction using petroleum ether (4060oC, BP) on a Soxtec System HT6 (Tecator application note 67/83). Crude fat in liver and carcass was
determined by the above procedure but without the acid hydrolysis. Crude fat in flesh was determined
by extraction in chloroform/methanol (2:1 v/v) as described by Folch et al. (1957).
Fibre; Crude fibre was determined by acidic followed by alkaline digestion and incineration of the dried
sample for 2h at 550oC.
Elemental analysis; Accurately weighed dry samples were subjected to wet-decomposition at 105oC
in 10 ml of concentrated nitric acid (Aristar, 16M) and allowed to cool. If the samples were not clear 3
ml of hydrogen peroxide (30% w/v) was added and the samples digested further at 105 oC. After
decomposition, samples were made up to 25 ml with purified, de-ionised water. Calcium and zinc
were then determined by carbon arc atomic absorption spectroscopy. Yttrium was determined using
inductively coupled plasma optical emission spectrophotometry (Thermo Jarell-Ash IRIS Axial ICPOES).
Phosphorous; Phosphorous was measured as inorganic phosphate following oxidation using acid
peroxide as described above and using the method of Teitz (1986). Phosphate forms a complex with
molybdenum and vanadate which has an absorption maximum at 405 nm.
Starch; Starch was assayed using an assay kit (SA-20) supplied by Sigma-Aldrich, (Poole, England).
Starch was hydrolysed by amyloglucosidase and glucose was phosphorylated by ATP in a reaction
catalysed by hexokinase. Glucose-6-P is then oxidised to 6-phosphoglucanate in the presence of NAD
in a reaction catalysed by glucose-6-P dehydrogenase During this reaction equimolar amounts of NAD
are converted to NADH and this can be followed by an increase in absorbance at 340 nm which is
directly proportional to glucose concentration.
Fatty acid analysis; Total lipid content of diet and tissue samples was determined gravimetrically after
extraction by homogenization in chloroform/methanol (2:1, v/v), basically according to Folch et al.
SID 5 (2/05)
Page 4 of 16
(1957). Fatty acid methyl esters (FAME) were prepared from total lipids by acid-catalyzed
transesterification in 2 ml of 1% H2SO4 in methanol plus 1 ml toluene as described by Christie (1982)
and FAME extracted and purified as described previously (Ghioni et al., 1996). FAME were separated
and quantified by gas-liquid chromatography (Carlo Erba Vega 8160, Milan, Italy) using a 30 m x 0.32
mm i.d. x 0.25 µm capillary column (CP wax 52CB; Chrompak Ltd., London, U.K). Hydrogen was used
as carrier gas and temperature programming was from 50oC to 150oC at 40oC/min and then to 230oC at
2oC/min. Individual methyl esters were identified by comparison to known standards and by reference
to published data (Ackman, 1980). Data were collected and processed using the Chromcard for
Windows (version 1.19) computer package (Thermoquest Italia S.p.A., Milan, Italy).
Definitions and calculation of indices
Digestibility and retention efficiency definitions
Digestibility = ADC = 100 - (100 * I in diet * N in faeces / I in faeces /
N in diet), where I = indicator and N = nutrient
Protein retention efficiency = PRE = ((P2 * W2)-(P1 * W1))/(FP * FI)*100,
where W is weight (g), P is protein concentration in fish (%), FP is
protein concentration in feed (%) and FI is feed intake
Energy retention efficiency = ERE = ((E2 * W2)-(E1 * W1))/(FE * FI)*100,
where W is weight (g), E is energy concentration in fish (MJ/kg), FE is
energy concentration in feed (MJ/kg) and FI is feed intake
Histology
30 pots of preserved tissues, containing anterior and posterior stomach, pyloric caecae/pancreas,
anterior gut, mid-gut, heart, liver, posterior stomach and hindgut, were examined histologically. Diet
type identification information was not supplied with samples so that histological examination was
carried out blind. 24 samples were taken comprising duplicate fish samples taken from 6 different
tanks.
4. Results
4.1 Digestibility
The apparent digestibility coefficients (ADCs) observed in the trial are presented in Table 1. One feed
sample and one faeces sample (with 3 samples pooled) were analysed per treatment. The faeces from
fish fed diet 4 (36% fullfat Soya) was mixed incorrectly, thus ADCs could not be calculated for this
treatment.
The ADCs for fat, protein and starch seemed to decrease with increasing levels of dietary fullfat Soya,
but a closer inspection of the results indicated a general ADC reduction as a result of fullfat Soya
inclusion and irrespective of fullfat Soya level (Figure 1). ADCphosphorous equaled zero when fullfat soya
was included in the diet.
SID 5 (2/05)
Page 5 of 16
Table 1: Apparent digestibility coefficients (ADC) for the main nutrients, energy, phosphorous and
selected fatty acids observed in the trial.
Level of fullfat soya (%)
Fat
Protein
Starch
Ash
NFE
GE
P
Tot sat
Tot mono
18:2n-6
Tot n-6
20:5n-3
22:6n-3
Tot n-3
0
93.7
90.6
89.0
44.8
55.7
87.1
27.3
65.4
80.1
82.0
79.7
97.6
95.5
96.8
12
87.7
85.2
84.7
23.2
49.1
81.2
0.6
54.2
74.1
77.6
76.8
94.8
93.0
93.1
24
86.6
86.0
83.6
20.9
42.3
80.7
2.9
52.0
72.4
80.7
80.5
95.1
93.3
93.2
36
100,0
95,0
Apparent digestibility (%)
90,0
y = -0,2983x + 92,908
R2 = 0,8678
Fat
Protein
Starch
85,0
y = -0,2217x + 88,436
R2 = 0,897
y = -0,1923x + 89,595
R2 = 0,6253
80,0
75,0
70,0
0
5
10
15
20
25
30
35
40
Inclusion level of fullfat soya (%)
Figure 1: ADCs for fat, protein and starch decrease with inclusion of fullfat soya in the diet, but
increasing levels do not seem to affect the ADCs further.
SID 5 (2/05)
Page 6 of 16
4.2 Bone mineralisation
Only the Zinc content of the bones was affected by dietary treatment. The bone Zn content increased
significantly with increasing levels of fullfat soya in the diet (Table 2).
Table 2: Bone minerals. Numbers are mean  SD of 3 replicate measurements per treatment.
Level of fullfat soya (%)
Zn
P
Ca
Ca/P ratio
0
763a
1423
20711
1.460.09
12
883ab
1534
20914
1.360.07
24
1018b
1517
20411
1.360.13
36
9513b
1503
2113
1.410.03
p
0.021
0.055
0.865
0.480
4.3 Whole body analysis
Inclusion of fullfat Soya did not have a marked effect on the whole body composition (Table 3). Neither
did inclusion of fullfat Soya at the levels tested in this trial seem to affect the whole body mineral
nutrition. This is contrary to what was expected due to the phytate content of the soya, which binds
minerals and make them unavailable to the animal. It should, however, be noted that the diets were
supplemented with additional calcium phosphate at increasing Soya inclusion levels, but other minerals
were supplied at the same level of supplementation.
Both protein and energy retention efficiencies were reduced by inclusion of fullfat Soya in the diet, but
they did not seem to be affected by the level of fullfat Soya (Figure 2).
Table 3: Whole body main nutrients and minerals. Numbers are mean  SD of 3 replicate samples per
treatment.
Level of fullfat soya (%)
Dry matter (%)
Ash (%)
Lipid (%)
Protein (%)
Zn (mg/kg)
P (mg/kg)
Ca (mg/kg)
SID 5 (2/05)
Start
24.11.2
3.10.2
3.50.3
16.71.0
18.7
3849
7673
0
28.01.1
2.70.2
8.21.7
16.10.5
14.22.4
3795243
6807974
12
27.60.6
2.70.0
7.80.5
16.40.4
14.50.2
3948219
6766649
Page 7 of 16
24
26.20.6
2.80.1
6.00.8
16.40.4
14.51.5
3849265
7062381
36
26.91.1
2.80.2
6.9 1.1
16.40.3
15.21.3
390199
66621103
70,00
60,00
PRE or ERE (%)
50,00
40,00
Contr
12
24
36
30,00
20,00
10,00
0,00
PRE 1
PRE whole
ERE 1
ERE whole
Figure 2: Protein (PRE) and energy (ERE) retention efficiencies during period 1 and the whole trial.
Numbers are mean  SD of 3 replicate measurements per treatment (based on dry matter).
4.4 Fatty acid profiles
The liver FA profile was almost a mirror image of the dietary FA profile (Figure 3). Although the livers
from the fish fed the fishmeal and fish oil based control diet (0% fullfat Soya) exceeded the LA level
(linoleic acid) of a maximum of 3% set in the European Pharmacopoeia standard, livers were high in
EPA and DHA in all treatments.
30,0
25,0
% of FA in liver
20,0
18:2n-6
20:5n-3
22:6n-3
15,0
10,0
5,0
max 3% 18:2 n-6
0,0
0,00
5,00
10,00
15,00
20,00
25,00
30,00
% of FA in diet
Figure 3: Dietary 18:2 n-6 (LA), 20:5 n-3 (EPA) and 22:6 n-3 (DHA) vs liver contents. Numbers are
means of duplicate analyses.
SID 5 (2/05)
Page 8 of 16
30,0
25,0
% of FA in muscle
20,0
18:2n-6
20:5n-3
22:6n-3
15,0
10,0
5,0
0,0
0,00
5,00
10,00
15,00
20,00
25,00
30,00
% of FA in diet
Figure 4: Dietary 18:2 n-6 (LA), 20:5 n-3 (EPA) and 22:6 n-3 (DHA) vs muscle contents. Numbers are
means of duplicate analyses.
EPA and DHA were selectively incorporated into the muscle, and incorporation of LA into the muscle
did not increase to the same extent as the increase in dietary content (Figure 4).
The fatty acid composition of muscle changed as the EPA and DHA content of the diet fell. The
differences were not significant. As muscle cells are normally of consistent composition in terms of total
lipid content and fatty acid makeup (Jangaard, Ackerman, Sipos, 1967), this unexpected finding
requires further investigation (Bob Ackerman, pers. comm., 2005).
4.5 Histology
A moderately significant inflammatory cell presence was noted in the posterior gut tissue from some of
the fish sampled that had been fed diet with 36% inclusion of fullfat soya (Appendix). Some had some
degree of posterior gut inflammation. Others only showed mild inflammatory cell numbers but also
showed other inflammatory cell foci in stomach lamina propria and sub-mucosal tissues. The severity of
the pathology seen and the numbers of associated inflammatory cells present even in the relatively
worst-affected fish appeared to be significantly less than those seen in Atlantic salmon fed similarly
substituted vegetable diets. However, these effects may affect performance following diet substitution
and the effects of fullfat Soya on the immune system need to be examined further.
5. Discussion and conclusions
The report presented here relates to proximal examination and other laboratory analyses and should
be considered together with report FC0930, “Substitution of fish meal with vegetable protein in cod
SID 5 (2/05)
Page 9 of 16
diets”. That report described growth and food conversion rate and although growth was not significantly
affected by the level of full fat Soya substitution over the full 12 week trial, the FCR was significantly
less in fish on 36% substituted diet compared with the control group.
The present study indicates that the effect of inclusion of Soya in the diet has an effect on digestibility
and whole body analysis but at the levels tested the quantity of Soya substituted, increasing the level
had no significant effect. This is also true of the growth rate and FCR of the fish. However, a cost
benefit analysis should be undertaken of the effect of substitution. Although growth rates may be
slightly lower the lower costs of ingredients may make substituted diets cost-effective.
The low levels of ash and phosphorus digestibility in the substituted diets indicate that the phytate in
the Soya, one of the important antinutritional factors in Soya which, binds with certain minerals, making
them difficult to absorb, was producing some effect on mineral availablity, but this had only a marginal
effect on mineral status of fish at the end of the trial.
The greatest effect of substitution was apparent in the fatty acid profiles. When the fatty acid profiles in
the diets and in the liver were compared EPA and DHA values were similar but the levels of linoleic
acid in the liver were higher. This would indicate that the livers could not be used for cod liver oil and
this should be considered when examining the cost of cod production when using the whole fish. The
livers could be used for other purposes such as pet foods and margarine.
The overall conclusion from the study is that cod can utilise the substituted raw materials. There is a
slight effect of inclusion of these raw materials on cod performance. However, a cost benefit analysis
should be undertaken of the value of including raw materials, such as whole soya protein, in cod diets.
6. References
Ackman, R.G., 1980. Fish lipids, part 1. In: (Connell, J.J. ed.) Advances in Fish Sciences and
Technology, Fishing News Books Ltd., Farnham, U.K. pp. 86-103.
AOAC 1995. Official Methods of Analysis of the Association of Offical Analytical Chemists International,
16th edn. Association of Official Analytical Chemists, Arlington, VA, USA.
Christie, W.W., 1982. Lipid Analyses, 2nd ed., Pergamon Press, Oxford, England. pp. 52-56.
Folch, J., Lees, M., Sloane-Stanley, G.H., 1957. A simple method for the isolation and purification of
total lipids from animal tissues. J. Biol. Chem. 226, 497-509.
Ghioni, C., Bell, J.G., Sargent, J.R., 1996. Polyunsaturated fatty acids in neutral lipids and
phospholipids of some freshwater insects. Comp. Biochem. Physiol. 114B, 161-170.
Jangaard, P.M., Ackerman, R.G., Sipos, J.C., 1967. Seasonal changes in fatty acid composition of cod
liver, flesh, roe and milt lipids. J. Fish. Res. Bd Can. 24, 613-627.
Teitz, W. 1986. Text Book of Clinical Chemistry, W.B. Saunders Inc. Philadelphia.
SID 5 (2/05)
Page 10 of 16
Appendix
Detailed histology reports by David Cox, Fish Vet Group
Key:
Control, normal marine meal and oil diet, no substitution= tanks N3, N7, N11
36% substitution with fullfat soya= tanks N5, N10, N13
CONTROL
N3 1A
Heart: OK
Stomach-transitional anterior to posterior: digesta present but otherwise OK
Anterior gut:autolytic loss of mucosal folds only, otherwise OK
N3 1B
Anterior stomach: OK
Pyloric caecae/pancreas: autolytic and artefactual type changes only
Intestine-multiple, convoluted mucosal folds overlying a very loosely-associated sub-mucosal
layer–? not posterior as pancreatic tissue attached: significant spread of mucous cells** plus a
mild, diffuse, patchy increased cellularity in sub-mucosal layer
** high numbers of mucous cells are commonly seen in the anterior gut in salmon-not clear about cod
Liver: OK
N3 2A
Heart: OK
Stomach-transitional anterior to posterior: digesta present but otherwise OK
Anterior gut: OK
N3 2B
Anterior stomach: digesta, occasional small area of inflammatory cells in the sub-mucosae otherwise
OK
Intestine –? posterior but pancreatic tissue attached: significant spread of mucous cells only, no
increased cellularity
Pyloric caecae/pancreas: autolytic and artefactual type changes plus occasional area of inflammatory
cells in lamina propria of mucosal folds
Liver: OK
N7 1A
Heart: OK
Stomach-transitional anterior to posterior: digesta present but otherwise OK
Anterior gut: OK
N7 1B
Pyloric caecae/pancreas: autolytic type changes only
Anterior stomach: OK
Intestine –? posterior but pancreatic tissue attached: significant spread of mucous cells only plus a
mild, diffuse patchy increased cellularity in sub-mucosal layer
Liver: OK
N7 2A
Heart: OK
Stomach-transitional anterior to posterior: digesta present but otherwise OK
Anterior gut: OK
SID 5 (2/05)
Page 11 of 16
N7 2B
Pyloric caecae/pancreas: autolytic type changes only
Anterior stomach: OK
Intestine –? posterior but pancreatic tissue attached: significant spread of mucous cells only
Liver: Large area with multiple small foci of scar tissue interspersed with inflammatory cells –around
bile ducts and around liver cells per se
N11 1A
Heart: OK
Posterior stomach: OK
Anterior gut: OK
N11 1B
Pyloric caecae/pancreas: Significant autolysis but otherwise OK; sub-mucosae very looselyconstructed!
Anterior stomach: OK
Intestine –? posterior but pancreatic tissue attached: significant spread of mucous cells, strands of
mucus and degenerate cellular material in gut lumen, ? increased connective tissue cells in submucosal layer
Liver: Moderate spread of recent and old haemorrhages surrounded by a chronic inflammatory
response
N11 2A
Heart: OK
Posterior stomach: OK
Anterior gut: Autolytic changes otherwise OK
N11 2B
Anterior stomach: OK
Pyloric caecae/pancreas: OK
Intestine –? posterior but pancreatic tissue attached: significant spread of mucous cells, OK
Liver: as N11 1B but fewer haemorrhages
36% SUBSTITUTION
N5 1A
Heart: a few, small areas of muscle inflammation
Posterior stomach: Large chronic inflammatory lesion in the sub-mucosal layer
Anterior gut: mucosal layer missing due to artefactual loss
N5 1B
Anterior stomach: OK, digesta present
Pyloric caecae/pancreas: Sub-mucosal layer very loosely associated,
Intestine – ? posterior but pancreatic tissue attached: significant spread of mucous cells,
significant accumulations of bacterial rods attached to mucosal cells and associated with strands of
mucus and other amorphous material. Mucosal folds very eosinophilic with loss of cell to cell border ?
incipient necrosis ? autolysis only
Liver: Minor spread of scar type lesions
N5 2A
Heart: solitary small area of muscle inflammation
Posterior stomach: OK
Anterior gut: autolytic type changes in mucosal folds only
N5 2B
Anterior stomach: OK, digesta present
Pyloric caecae/pancreas: Sub-mucosal layer very loosely associated, autolytic changes
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Intestine – ? posterior but pancreatic tissue attached: autolytic changes, significant spread of
mucous cells, otherwise OK
Liver: OK
N10 1A
Heart: a few, small areas of muscle inflammation
Posterior stomach: diffuse increase in numbers of connective tissue cells in sub-mucosal layer
Gut: autolytic type changes in mucosal folds only
N10 1B
Anterior stomach: Epithelial cells at the tips of the mucosal folds appear to have lost their definition,
folds in general look crypt-like as if shrunken!
Pyloric caecae/pancreas: Significant autolytic type changes affecting mucosal folds, patchy increased
cellularity in lamina propria, sub-mucosal layer very loosely associated
Intestine – ? posterior but pancreatic tissue attached: significant spread of mucous cells, appears
to be a significantly increased cellularity in the lamina propria and the sub-mucosal layer but large
areas of the mucosal epithelium was missing!
Liver: OK
N10 2A
Heart: a few, small areas of muscle inflammation
Posterior stomach: diffuse increase in numbers of connective tissue cells in sub-mucosal layer plus
patchy inflammatory cells in lamina propria in mucosal folds
Gut: Loss of mucosal epithelial layer and the sub-mucosal layer appeared to have higher numbers of
connective tissue cells
N10 2B
Anterior stomach: OK
Pyloric caecae/pancreas: significant autolysis, patchy increased cellularity in lamina propria plus submucosal layer very loosely associated
Intestine – ? posterior but pancreatic tissue attached: significant spread of mucous cells, significant
patchy increase in cellularity in lamina propria, sub-mucosal layer very loosely associated, considerable
digesta with accumulations of bacterial rods
Liver: OK
N13 1A
Heart: OK
Stomach-transitional anterior to posterior: moderate, patchy increase in cellularity in the lamina
propria and the sub-mucosae
Gut: Significant autolytic changes with loss of mucosal fold epithelia, mild, patchy increased cellularity
in lamina propria plus sub-mucosal layer very loosely associated
N13 1B
Anterior stomach: OK
Pyloric caecae/pancreas: significant autolysis
Intestine – ? posterior but pancreatic tissue attached: significant spread of mucous cells, autolytic
changes, sub-mucosal layer very loosely associated,
Liver: minor, patchy granulomatous haemorrhages
N13 2A
Heart: OK
Posterior stomach: patchy increase in cellularity in the lamina propria and the sub-mucosae
Gut: Loss of mucosal epithelial layer and the sub-mucosal layer appeared to have higher numbers of
connective tissue cells
N13 2B
Anterior stomach: moderately increased cellularity in sub-mucosal layer
Pyloric caecae/pancreas: patchy increase in cellularity in lamina propria plus sub-mucosal layer very
loosely associated,
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Intestine – ? posterior but pancreatic tissue attached: significant spread of mucous cells, autolytic
changes, significant, patchy increased cellularity in lamina propria, sub-mucosal layer very loosely
associated
Liver: minor, patchy granulomatous haemorrhages
Sub 1A
Pyloric caecae/pancreas: significant autolytic changes, significant increase in cellularity in the lamina
propria, sub-mucosal layer very loosely associated
Anterior stomach: OK
Sub 1B
Heart: OK
Intestine– ? posterior but pancreatic tissue attached: autolytic changes, patchy increase in
cellularity in lamina propria, sub-mucosal layer very loosely associated
Posterior stomach: OK
Sub 2A
Pyloric caecae/pancreas: marked increased cellularity in lamina propria, sub-mucosal layer very
loosely associated
Anterior stomach: very little in section
Intestine– ? posterior but pancreatic tissue attached: significant spread of mucous cells, significant
autolytic changes, patchy increase in cellularity in lamina propria, sub-mucosal layer very loosely
associated
Sub 2B
Heart: OK
Intestine– ? posterior but pancreatic tissue attached: significant spread of mucous cells, significant
autolytic changes, no other findings
Posterior stomach: OK
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References to published material
9.
This section should be used to record links (hypertext links where possible) or references to other
published material generated by, or relating to this project.
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Related report:
DEFRA report FC0930, project ”Substitution of fish meal with vegetable protein on cod diets.”
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