Report Confidential K-300 (cid:121) Published 23 December 2005
`
`F/T ATLANTIC NAVIGATOR 2004 -2005
`
`Eyolf Langmyhr
`
`AKER EXHIBIT 2003 PAGE 0001
`
`

`

`Norut Group Ltd. consists of six research
`institutes located in Tromsø, Narvik and Alta.
`The Norut Group has 220 employees whose
`applied research and development
`encompasses a wide variety of
`interdisciplinary fields. Each subsidiary
`institute has a specific research emphasis,
`but common to all is activity centered
`around the polar and Barents regions.
`
`Norut Group LTD consist of:
`
`Fiskeriforskning (Norwegian Institute of
`Fisheries and Aquaculture Research), Tromsø
`and Bergen
`
`Norut Information Technology Ltd, Tromsø
`
`Norut Social Science Researc Ltd, Tromsø
`
`Norut Technology Ltd, Narvik
`
`Norut Medicine and Health Ltd, Tromsø
`
`Norut NIBR Finnmark AS, Alta
`
`Fiskeriforskning (Norwegian Institute of
`Fisheries and Aquaculture Research) conducts
`research and development for the fisheries and
`aquaculture industry. The Institute covers
`virtually all links in the value chain – “from sea
`bottom to tabletop”. Fiskeriforskning is a
`national research institute – owned by the
`Norut Group Ltd. (51 %) and the Norwegian
`Ministry of Fisheries (49 %). Located in
`Tromsø (head office) and Bergen, the facilities
`at Fiskeriforskning are an important part of the
`national infrastructure for fisheries and
`aquaculture research.
`
`Fiskeriforskning have five main areas of
`research:
`(cid:130) Seafood and industrial processing
`(cid:130) Marine biotechnology and fish health
`(cid:130) Aquaculture
`(cid:130) Aquafeed and marine processing
`(cid:130)
`Economics and marketing
`
`Tromsø (head office)
`Muninbakken 9-13, Breivika
`P.O.box 6122
`NO-9291 Tromsø
`Norway
`Tel.: +47 77 62 90 00
`Fax: +47 77 62 91 00
`E-mail:post@fiskeriforskning.no
`
`Bergen
`Kjerreidviken 16
`NO-5141 Fyllingsdalen
`Norway
`Tel.: +47 55 50 12 00
`Fax: +47 55 50 12 99
`E-mail: office@fiskeriforskning.no
`
`Internet: www.fiskeriforskning.no
`
`AKER EXHIBIT 2003 PAGE 0002
`
`

`

`Norwegian Institute of Fisheries and
`Aquaculture Research
`Main office: P.O.box 6122, NO-9291 Tromsø
`Visiting address: Muninbakken 9-13,
`Tlf.: +47 77 62 90 00, fax: +47 77 62 91 00
`E-mail: post@fiskeriforskning.no
`
`Dept. Bergen: Kjerreidviken 16, NO-5141 Fyllingsdalen
`Tlf.: +47 55 50 12 00, fax: +47 55 50 12 99
`E-mail: office@fiskeriforskning.no
`
`Internet: www.fiskeriforskning.no
`
`Organisation no.: NO 964 441 898 MVA
`
`Accessibility:
`Confidential
`
`Report K-300
`
`Date:
`23 December 2005
`Number of pages and appendixes:
`25
`Director of Research:
`Ola Flesland
`Project no:
`5560
`Employers ref.:
`Finn-Arne Lorentsen
`
`REPORT
`
`Tittel:
`F/T ATLANTIC NAVIGATOR 2004 -2005
`
`Author (s):
`Eyolf Langmyhr
`By agreement with:
`Aker Seafoods
`3 keywords:
`Krill, krillmeal, krilloil
`Summary:
`The report is a collection of all analyses carried out at Fiskeriforskning Analytical Laboratory in
`2004 and 2005 on samples from F/T Atlantic Navigator. The analytical results are sorted and
`evaluated.
`The microbial standard of the krill meal is good.
`The moisture content varies between 6.0 and 7.8 g/100 g, which is a satisfactory range. Too dry
`and the oxidation stability are reduced. Too wet and fungus may grow.
`The variation in the fat content of the krill meal follows the seasonal variation of the fat content of
`krill raw material. The process is not capable of adjusting for the variation in the fresh krill.
`The high proportion of phospholipids in krill (40 %) makes it difficult to separate the fat by only
`heat and mechanical treatment.
`Most of the oxidation reactions in the process take place in the dryer.
`The flow properties of krill meal are good for meals with less than 18 % fat.
`The Kett analyser gives high values for protein, low for ash, and lower for fluorine then the
`chemical analyses. The results for moisture, fat and salt are in agreement with the chemical
`analyses. With a proper calibration this instrument should work well for process control.
`
`AKER EXHIBIT 2003 PAGE 0003
`
`

`

`TABLE OF CONTENTS
`
`1
`2
`
`3
`
`4
`
`INTRODUCTION ............................................................................................................ 1
`ANALYTICAL METODS ............................................................................................... 1
`2.1 Method references ................................................................................................... 1
`2.2 Abbreviations .......................................................................................................... 2
`2.3 Presentation of the results........................................................................................ 2
`MICROBIOLOGICAL ANALYSES............................................................................... 3
`3.1 Frozen krill .............................................................................................................. 3
`3.2 Krill meal................................................................................................................. 4
`3.3 Krill oil .................................................................................................................... 4
`COMPOSITION AND QUALITY OF MEAL ................................................................ 5
`4.1 Major constituents ................................................................................................... 5
`4.2 Minerals and quality parameters ............................................................................. 8
`4.3 Astaxanthin.............................................................................................................. 9
`4.4 Antioxidant............................................................................................................ 10
`4.5 Amino acids........................................................................................................... 11
`4.6 Fatty acid distribution............................................................................................ 12
`4.7 Physical characteristics.......................................................................................... 13
`4.8 Kett Composition Meter........................................................................................ 14
`4.8.1 Kett analyses 2005 ....................................................................................... 15
`KRILL OIL..................................................................................................................... 16
`PROCESS ....................................................................................................................... 18
`6.1 Process samples..................................................................................................... 18
`6.2 Evaluation of the krill processing line................................................................... 18
`6.3 Mince test .............................................................................................................. 21
`APPENDIX .............................................................................................................................. 22
`Table 18. Composition of krill meal...................................................................... 22
`Table 19. Astaxanthin in krill meal ....................................................................... 24
`Table 20. Ethoxyquin in krill meal ........................................................................ 25
`
`5
`6
`
`AKER EXHIBIT 2003 PAGE 0004
`
`

`

`1
`INTRODUCTION
`By request from Finn-Arne Lorentsen, Aker Seafoods, all analyses carried out at
`Fiskeriforskning Analytical Laboratory in 2004 and 2005 on samples from F/T Atlantic
`Navigator are collected. In this report the analytical results are sorted and evaluated. The
`evaluation is based on production reports from Atlantic Navigator and advices and
`discussion on processing.
`
`2
`
`ANALYTICAL METODS
`
`2.1 Method references
`Protein, Kjeldahl’s method: ISO 5983:1997, Method A 01. Uncertainty: 1 %.
`
`Nitrogen, Combustion: ISO/DIS 16634, Method A 25.
`
`Water Soluble Crude Protein: In house method A 20.
`
`Digestible protein, Mink: Skrede A., Acta Agric. Scandinavica, 29, 241-257, 1979.
`Mundheim H. og Opstvedt J., The current status of fish nutrition in Aquaculture,
`Laboratory of fish nutrition, Tokyo, Japan, 195-200, 1989. M. Takedo and T. Watzuake
`(Editors). Method A 24.
`
`Moisture/Dry matter: ISO 6496 (1999). Method A 04. Uncertainty: 4 %.
`
`Ash: ISO 5984:2002. Method A 02. Uncertainty: 3 %.
`
`Fat, Ethyl acetat extraction: NS 9402, 1994 (modified calculation). Method A 29.
`
`Fat, Soxhlet: AOCS Official Method Ba 3-38 Reapproved 1993. Method A 03.
`
`Fat, Acid hydrolysis: Commission Directive 98/64/EC. Method A38.
`
`Fat, Bligh and Dyer: E.G. Bligh & W.J. Dyer: A rapid method of total lipid extraction and
`purification. Can.J.Biochem.Physiol. Vol 37 (1959). Methode A 56.
`
`Salt (NaCl): AOAC, Methods of analysis, 16th edition, 937.09. Method A 06.
`
`Amino acids: Waters Accq-Tag Amino Acid Analysis Method. Analytical Biochemistry
`211, 279-287 (1993). Method A 42.
`
`Ethoxyquin, Fluorimetrisk metode: AOAC Official Method 963.07, Modification: Hexan
`replaces petroleums ether, pkt. 5. C. Method A 44.
`
`Flow number: Nilsen, Ø. Hjelpestoffer - et alternativ for å bedre flyt-egenskapene til
`sildemel. SSF Report B337 (1981). Method A35.
`
`Astaxanthin: Schierle J. & Härdi W. 1994. Determination of stabilized astaxanthin in
`Carophyll® Pink, premixes and fish feeds. Edition 3. Revised Supplement to: Hoffman P,
`
`1
`
`AKER EXHIBIT 2003 PAGE 0005
`
`

`

`Keller HE, Schierle J., Schuep W. Analytical methods for vitamins and carotenoids in feed.
`Basel: Department of Vitamin Research and Development, Roche. Method A 23.
`
`Fatty acid distribution: AOCS Official Method Ce 1b-89. Method A 68.
`
`Moisture in oil: AOCS Official Method CA 2e-84. Reapproved 1993. Method A 13.
`
`Unsaponifiable Matter: AOCS Official Method Ca 6b-53 Reapproved 1993. Method A 08.
`
`Free Fatty Acids: AOCS Official Method Ca 5a-40 Reapproved 1993. Method A 07.
`
`Anisidine Value: AOCS Official Method Cd 18-90 Reapproved 1993. Method A 09.
`
`Peroxide Value: AOCS Official Method Cd 8b-90 Official 1996. Method A 10
`
`Aerobic Plate Count: NMKL No. 146, 1993. ISO 6887, 1983. Petrifilm, 3M – avlesnings-
`mal. Method A 16.
`
`Sulphite-reducing clostridia: NMKL no. 56, 1994. Method A 27.
`
`E. coli and Coliform bacteria: NMKL No 147, 1993. Modifocations: ISO 6887-1-1999
`(dilution medium), ISO 7218-1996 w. amendment 1-2001(calculation), Avläsningsmall
`Petrifilm for coliforme bakterier og E.coli (3M). Method A 46.
`
`Listeria monocytogenes: NMKL No. 136, 2. utg., 1999. Method A 70.
`
`Salmonella Bacteria: AOAC Official Method 991.38, 16th ed. 1996, NMKL No. 71, 1991,
`Method A17.
`
`Enterobacteriaceae: ISO 7402, 1985. Method A19.
`
`H2S producing bacteria: NMKL No. 96. Method A16.
`
`Aerobic plate count at 20 °C: NMKL No. 96.
`
`2.2 Abbreviations
`AOAC - AOAC INTERNATIONAL
`
`AOCS - The American Oil Chemists' Society
`
`ISO - International Organization for Standardization
`
`NMKL - Nordisk Metodikkomité for Næringsmidler
`
`2.3 Presentation of the results
`An empty space in the table means that the component has not been analysed.
`
`< means that the concentration of the component is below the given limit of detection.
`
`mg/kg is the same as ppm, g/100 g is the same as %, mg/kg = % × 10000.
`
`2
`
`AKER EXHIBIT 2003 PAGE 0006
`
`

`

`3 MICROBIOLOGICAL ANALYSES
`
`3.1 Frozen krill
`Two samples of frozen krill are analysed to get information on bacterial count in the raw
`krill. The parameters give information on total bacterial count (aerobic plate count:
`bacteria that grow in the presence of air/oxygen, and sulphite reducing clostridia: bacteria
`that only grow in the absence of oxygen). The aerobic plate count was determined by a
`method dedicated to fish (20 °C) and the standard method which is planned used in the
`production control on Saga Sea. Hydrogen sulphide producing bacteria (mainly
`Shewanella putrefaciens) are considered among the most important quality deteriorating
`bacteria in seafood. The presence of Enterobacteriaceae is normally an indication of faecal
`contamination, but may also be present in the natural bacterial flora of seafood.
`Enterobacteriaceae produce repulsive odour due to reduction of trimethylamine oxide. The
`requirement of a maximum allowed count of Enterobacteriaceae is based on the indication
`of faecal contamination. The results in Table 1 show that the samples have an acceptable
`total bacterial load, and the load of the most active quality deteriorating bacteria is small.
`
`Table 1. Bacteria in round frozen krill from Atlantic Navigator 27.02.05
`The samples were taken from two different boxes.
`
`per gram
`Aerobic plate count
`per gram
`H2S producing bacteria
`per gram
`Enterobacteriaceae
`per gram
`Sulphite reducing clostridia
`per gram
`Aerobic plate count at 20 °C
`Aerobic plate count was determined by two methods:
`Standard method which are planned used in production control on Saga Sea
`Method dedicated to fish (20 °C)
`
`Box 1
`78000
`150
`<10
`<10
`100000
`
`Box 2
`700
`<10
`<10
`170
`530
`
`3
`
`AKER EXHIBIT 2003 PAGE 0007
`
`

`

`3.2 Krill meal
`Microbiological analyses were performed in two instances. For the first productions the
`aim was to ensure that no harmful bacteria were present in the meal. Later the general
`microbial load was examined.
`
`The microbial quality of the krill meals was satisfactory. No harmful bacteria were
`detected.
`
`Table 2. Bacteria in krill meal
`Sulphite
`Aerobic
`reducing
`plate
`E. coli
`clostridia
`count
`per gram per gram per gram
`<10
`<10
`<10
`<10
`<10
`<10
`<10
`<10
`<10
`<10
`<10
`
`Date
`02.01.04
`04.01.04
`09.01.04
`12.01.04
`16.01.04
`20.01.04
`21.01.04
`23.01.04
`24.01.04
`27.01.04
`28.01.04
`28.01.-06.02.05
`28.01.05
`02.02.05
`09.02.05
`16.02.05
`21.02.05
`06.05.05
`Jul 10 - Jul 29
`Jul 30 - Aug 15
`May 28 - Jun 19
`Jun 29 - Jul 5
`
`5500
`<2500
`<2500
`<2500
`<2500
`<2500
`
`25000
`<2500
`5000
`2600
`
`<10
`<10
`<10
`<10
`<10
`<10
`
`<10
`<10
`<10
`<10
`
`Coliform. bact.
`at 37°C
`per gram
`<10
`<10
`<10
`<10
`<10
`<10
`<10
`<10
`<10
`<10
`<10
`
`Listeria monocyt.
`Salmonella negativ
`negative
`samples á 25 gram samples á 25 gram
`1
`1
`1
`1
`1
`1
`1
`1
`1
`1
`1
`1
`1
`1
`1
`1
`1
`1
`1
`1
`1
`1
`
`3
`
`From the meal produced 6.5.05 230 bags of krill meal was stored at Fiskeriforskning,
`Kjerreidviken, for research purpose. The salmonella analyses is analysed in a composite
`sample from 23 bags.
`
`3.3 Krill oil
`No bacteria were detected in the krill oil.
`
`Table 3. Microbiology of krill oil received 23.06.2004
`
`per gram
`Total plate count
`Coliform bacteria at 37°C per gram
`Salmonella not detected
`samples à 25 gram
`
`<2500
`<10
`1
`
`4
`
`AKER EXHIBIT 2003 PAGE 0008
`
`

`

`4
`
`COMPOSITION AND QUALITY OF MEAL
`
`4.1 Major constituents
`The analytical results are found in Table 18 in the appendix. For the first meal samples fat
`was determined by the Soxhlet method. This method gives too low results for krill meal,
`and a step with acid hydrolysis was added. Bligh and Dyer extract is used for analyses of
`the lipid phase.
`
`Protein varies between 55.0 and 66.9 g/100 g (Figure 1 and Table 18). In 2005 the protein
`content increases from 56 g/100 g in February to 64 g/100 g in August. The fat content of
`the krill meal varies between 8.4 and 22.3 g/100 g in 2004, and the seasonal variation can
`not be distinguished from changes in processing conditions. In 2005 the fat content went
`from 26 g/100 g in March to 13 g/100 g in August.
`
`In the first productions the moisture in the krill meal showed great variation, but the
`moisture content soon stabilised between 5.4 and 8.7 g/100 g for the rest of 2004, except
`for one sample with 3.5 g/100 g moisture. In 2005 the moisture varies between 6.0 and 7.8
`g/100 g. A good range is 6 – 9 g/100 g moisture. Too dry and the oxidation stability are
`reduced. Too wet and fungus may grow.
`
`75
`
`70
`
`65
`
`60
`
`55
`
`50
`
`Protein (g/100 g)
`
`Moisture
`Fat
`Protein
`
`45
`11.08.05
`
`30
`
`25
`
`20
`
`15
`
`10
`
`5
`
`Moisture, Fat (g/100 g)
`
`0
`01.01.04
`
`25.03.04
`
`17.06.04
`
`09.09.04
`
`02.12.04
`
`24.02.05
`
`19.05.05
`
`Date
`
`Figure 1. Protein, moisture and fat in krill meal
`Protein values on the right axis. The unit on the x-axis is 12 weeks. For composite samples the
`results are placed in the middle of the sampling period (Table 18).
`
`5
`
`AKER EXHIBIT 2003 PAGE 0009
`
`

`

`Ash
`
`Salt
`
`Saltfree ash
`
`20
`
`18
`
`16
`
`14
`
`12
`
`10
`
`02468
`
`g/100 g
`
`01.01.04
`
`25.03.04
`
`17.06.04
`
`09.09.04
`
`02.12.04
`
`24.02.05
`
`19.05.05
`
`11.08.05
`
`Figure 2. Ash and salt in krill meal
`
`Date
`
`The ash originates mainly from seawater and shell. The variation in ash is due to treatment
`of raw material and processing conditions. The variation of ash was 10.4 to 17.8 g/100 g in
`2004 and 10.3 to 12.6 g/100 g in 2005.
`
`The variation in salt content is small, 2.5 to 3.8 % in 2004 and 3.1 to 3.8 % in 2005.
`
`Figure 2 also shows saltfree ash (ash – salt) which sometimes are used in product data
`sheets for fish meal.
`
`3000
`
`2500
`
`2000
`
`1500
`
`mg/kg
`
`1000
`
`500
`
`0
`01.01.04
`
`25.03.04
`
`17.06.04
`
`09.09.04
`
`02.12.04
`
`24.02.05
`
`19.05.05
`
`11.08.05
`
`Figure 3. Fluorine in krill meal
`
`Date
`
`6
`
`AKER EXHIBIT 2003 PAGE 0010
`
`

`

`The content of fluorine seems to have stabilised on a lower level in 2005 than in 2004. The
`very high contents are found only in meal from the first productions. The variation was
`1100 to 2800 mg/kg in 2004 and 790 to 1700 mg/kg in 2005.
`
`By comparing Figure 3 with Figure 2 we see that the high fluorine contents are found in
`the meals with high ash content. By presuming that most of the salt comes from sea water,
`fluorine is plotted against saltfree ash. Fluorine in samples of meal from krill mince
`(Chapter 6.3) is included in Figure 4.
`
`3000
`
`2500
`
`2000
`
`1500
`
`1000
`
`Fluorine (mg/kg)
`
`500
`
`5
`
`6
`
`7
`
`Figure 4. Fluorine as a function of salt free ash.
`
`9
`8
`Saltfree ash (g/100 g)
`
`10
`
`11
`
`12
`
`The correlation between fluorine and saltfree ash in krill meal is described by regression
`(
`)
`Fluorine
`Ash
`Salt
`equation:
`.
`.0
`(%)
`0334
`127
`.0
`=
`×
`−
`−
`Standard error: 0.022. Correlation coefficient: 0.91.
`
`Fluorine
`For ash the regression gives:
`(%)
`.0
`0323
`=
`Standard error: 0.025. Correlation coefficient: 0.89.
`
`(
`
`×
`
`Ash
`
`−
`
`Salt
`
`)
`
`−
`
`.0
`
`228
`
`.
`
`As long as the saltfree ash is below 9.5% or the ash is below 13 % the fluorine content
`should be acceptable.
`
`7
`
`AKER EXHIBIT 2003 PAGE 0011
`
`

`

`4.2 Minerals and quality parameters
`
`Table 4. Minerals and quality parameters in krill meal
`
`Calcium
`
`Iron
`
`Magnesium Phosphorus
`
`NH3-N
`
`Cadaverine Histamine
`
`Date
`02.01.04
`04.01.04
`09.01.04
`12.01.04
`16.01.04
`21.01.04
`23.01.04
`24.01.04
`27.01.04
`28.01.04
`
`Min
`Max
`
`g/100 g mg/kg
`2.7
`151
`3.2
`51
`
`g/100 g
`0.77
`0.82
`
`3.0
`2.5
`3.0
`2.9
`2.9
`2.9
`3.0
`
`2.5
`3.2
`
`70
`44
`48
`69
`56
`57
`36
`
`36
`151
`
`0.73
`0.60
`0.73
`0.70
`0.77
`0.76
`0.75
`
`0.60
`0.82
`
`g/kg
`<0.1
`<0.1
`<0.1
`<0.1
`<0.1
`<0.1
`<0.1
`<0.1
`<0.1
`<0.1
`
`g/100 g
`1.56
`1.77
`1.77
`1.68
`
`1.55
`1.55
`
`1.56
`1.62
`
`1.55
`1.77
`
`g/100 g
`0.06
`0.04
`0.05
`0.04
`0.06
`0.04
`0.04
`0.04
`0.06
`0.05
`
`0.04
`0.06
`
`g/kg
`<0.01
`<0.01
`<0.01
`<0.01
`<0.01
`<0.01
`<0.01
`<0.01
`<0.01
`<0.01
`
`Peroxide value
`in Bl&D extract
`meq peroxide
`/kg lipid
`
`<1
`
`6.6
`
`<1
`<1
`6
`<1
`
`Minerals in krill meal was only analysed in the first produced meals. The quality
`parameters were good. Ammonia nitrogen is low, no biogenic amines (cadaverine and
`histamine) were detected, and the peroxide values are low.
`
`Biological digestible protein in krill meal was 85.4 g/100 g protein. The krill meal tested
`was composite sample from 11.03.-07.04.05, Trip 2. This is a rather low value for
`biological digestible protein, and krill protein is expected to be well digested. In addition to
`a possible reduced protein quality due to high temperature in the dryer, the chitin in the
`krill shell will have influence on the result for digestibility. Chitin composition of whole
`Antarctic krill is between 2.4 and 2.7 % of the dry weight. The nitrogen in chitin will be
`reported as protein, and if the chitin nitrogen is subtracted, the protein in meal from whole
`krill will be approximately 1 % lower. In meal without the water soluble proteins the effect
`of chitin will be greater, and in faeces from mink even greater. The effect of chitin on the
`analysis of biological digestible krill is not known, but if it had been accounted for, the
`digestible protein would be higher.
`
`8
`
`AKER EXHIBIT 2003 PAGE 0012
`
`

`

`4.3 Astaxanthin
`By excluding the first samples the variation in astaxanthin content is 99 – 156 mg/kg in
`2004 and 110 – 158 mg/kg in 2005. The price for synthetic astaxanthin is approximately kr
`15000 per kg, that is kr 1.50 per 100 mg. Astaxanthin is a fat soluble component but there
`is no relation between fat and astaxanthin in krill meal.
`
`160
`
`150
`
`140
`
`130
`
`120
`
`110
`
`100
`
`90
`
`80
`
`70
`
`mg/kg
`
`60
`01.01.04
`
`25.03.04
`
`17.06.04
`
`09.09.04
`
`Date
`
`02.12.04
`
`24.02.05
`
`19.05.05
`
`11.08.05
`
`Figure 5. Astaxanthin in krill meal
`
`Practically all of the astaxanthin in krill meal is in esterified (Table 19), and it is mostly
`esterified to two fatty acids. Synthetic astaxanthin, which are used in fish feed, are not
`esterified. Published results disagree on the efficacy of pigmentation by the esterified
`astaxanthin in krill meal compared to free astaxanthin.
`
`9
`
`AKER EXHIBIT 2003 PAGE 0013
`
`

`

`4.4 Antioxidant
`
`4000
`
`3800
`
`1600
`
`1400
`
`1200
`
`1000
`
`800
`
`600
`
`400
`
`200
`
`Ethoxyquin (mg/kg)
`
`0
`09.09.04
`
`21.10.04
`
`02.12.04
`
`13.01.05
`
`Figure 6. Ethoxyquin in krill meal
`The unit on the x-axis is 6 weeks.
`
`24.02.05
`Date
`
`07.04.05
`
`19.05.05
`
`30.06.05
`
`11.08.05
`
`In the first productions krill meal was stabilised with Seldox, a mixture of natural
`antioxidants and ethoxyquin (Selko, Nederland). Some of the krill meals had less
`ethoxyquin than required by IMO, 100 mg/kg. In 2005 Seldox was replaced by ethoxyquin.
`The concentration of ethoxyquin in the meal increased, but to unnecessary high levels
`(Figure 6). Towards the end of the season it seems that control with addition of ethoxyquin
`was improved and the concentration in the meal was stabilised at 300 - 400 mg/kg.
`
`Table 5. Ethoxyquin in stored krill meal
`Sample
`date
`20.04.05
`21.04.05
`22.04.05
`23.04.05
`27.04.05
`Mean
`20.04.05
`21.04.05
`22.04.05
`23.04.05
`27.04.05
`Mean
`
`Analysed
`24.06.05
`24.06.05
`24.06.05
`24.06.05
`24.06.05
`
`24.06.05
`24.06.05
`24.06.05
`24.06.05
`24.06.05
`
`mealroom
`meal room
`meal room
`meal room
`meal room
`
`cold room
`cold room
`cold room
`cold room
`cold room
`
`Ethoxyquin
`mg/kg
`160
`120
`130
`130
`210
`150
`220
`120
`170
`150
`190
`170
`
`Krill meal has been stored in the meal room and in a cold room on board Atlantic
`Navigator (Table 5). There seems to be no difference in shelf life between storage in meal
`room or cold room.
`
`10
`
`AKER EXHIBIT 2003 PAGE 0014
`
`

`

`4.5 Amino acids
`All teleost fish require 10 essential amino acids (EAA), namely, arginine, histidine,
`isoleucine, leucine, lysine, methionine, threonine, tryptophan, phenylalanine, valine (D. P.
`Bureau and C.Young Cho, http://www.uoguelph.ca/fishnutrition/feedint.shtml#1. Protein
`and Amino Acid Requirements of Fish). The requirement is shown together with amino
`acid composition in krill meal in Table 6. The sulphur containing amino acids methionine
`and cystein can be assessed together. The same goes for phenylalanine and tyrosine.
`
`The amino acid composition in krill meal is adequate as protein source for feed. The
`absence of hydroxyproline confirms that there is little connective tissue in krill.
`
`The water soluble protein in krill meal produced 06.06.2005 was 11.8 g/100 g protein. For
`Norse-LT 94 fish meal the water soluble protein is between 18 and 32 %. The content of
`the free amino acid taurin is 0.5 g/100 g protein (Table 6) while the content of taurin in
`fresh krill is reported to be about 2.8 g/100 g protein (T. Ellingsen, Biokjemiske studier
`over antarktisk krill, NTH 1982). The proportion of the free amino acids in that study is
`close to the values in Table 6.
`
`Table 6. Amino acids in krill meal and stickwater.
`The meal was produced 06.05.2005. The stickwater is from Trip 03.05 (See 6.1Process samples)
`
`Total
`g /100 g protein
`10.5
`13.5
`<0.5
`4.2
`4.4
`2.1
`6.7
`4.1
`5.4
`3.8
`4.0
`5.0
`2.9
`5.0
`7.8
`4.4
`7.8
`1.4
`1.1
`
`1.6
`4.2
`2.0
`
`2.2
`1.7
`2.0
`3.6
`2.7
`4.8
`
`0.6
`
`Amino acid
`Aspartic acid
`Glutamic acid
`Hydroxiproline
`Serine
`Glycine
`Histidine
`Arginine
`Threonine
`Alanine
`Proline
`Tyrosine
`Valine
`Methionine
`Isoleucine
`Leucine
`Phenylalanine
`Lysine
`Cysteine/Cystine
`Tryptophan
`Creatinine
`Asparagine
`Glutamine
`3-aminopropanoic acid
`Taurine
`4-aminobutanoic acid
`Citrulline
`Carnosine
`Anserine
`Ornithine
`3-aminopropanoic acid is also known as β-alanine
`4-aminobutanoic acid is also known as γ-aminobutyric acid or GABA
`
`Meal
`Requirements
`g /100 g protein
`
`(+ Cys 2.4)
`
`(+Tyr 5.3)
`
`11
`
`Free
`g /100 g protein
`0.02
`0.07
`<0.01
`0.02
`0.18
`<0.01
`0.56
`<0.01
`0.08
`0.53
`0.01
`0.02
`<0.01
`0.02
`0.14
`0.01
`0.02
`<0.01
`<0.02
`<0.01
`<0.01
`<0.01
`0.5
`0.5
`<0.01
`0.04
`<0.01
`<0.01
`0.02
`
`Stickwater
`Free
`g /100 g protein
`0.22
`0.51
`<0.05
`0.13
`3.28
`<0.05
`4.86
`0.22
`0.87
`2.32
`0.20
`0.13
`0.12
`0.1
`0.19
`0.10
`0.27
`<0.05
`<0.05
`<0.05
`0.05
`<0.05
`8.99
`8.52
`<0.05
`0.14
`<0.05
`<0.05
`1.04
`
`AKER EXHIBIT 2003 PAGE 0015
`
`

`

`4.6 Fatty acid distribution
`Fatty acid distribution is only analysed in the first productions. A sum of fatty acids of 60
`% is expected in a lipid with 40 % phospholipids and a few percent wax esters (Table 15).
`The lipid classes was analysed in separate study on evaluation of the processing line.
`
`The main fatty acids in the residual fat in krill meal is palmitic acid (16:0), oleic acid
`(18:1), EPA (ecosapentenoic acid, 20:5) and DHA (docosahexenoic acid, 22:6).
`
`Table 7. Fatty acid distribution in krill meal
`The first number is the number of carbon atoms in the fatty acid molecule. The number after the
`colon is the number of double bonds. The number after n is the place of the double bond furthest
`from the acid group.
`Fatty acid
`Date
`14:0
`g/100g lipid
`16:0
`g/100g lipid
`16:1 n-7
`g/100g lipid
`16:2 n-4
`g/100g lipid
`16:3 n-4
`g/100g lipid
`16:4 n-1
`g/100g lipid
`18:0
`g/100g lipid
`18:1 n-9+7+5
`g/100g lipid
`18:2 n-6
`g/100g lipid
`18:3 n-3
`g/100g lipid
`18:3 n-6
`g/100g lipid
`18:4 n-3
`g/100g lipid
`20:0
`g/100g lipid
`20:1 n-9+7
`g/100g lipid
`20:2 n-6
`g/100g lipid
`20:3 n-3
`g/100g lipid
`20:3 n-6
`g/100g lipid
`20:4 n-3
`g/100g lipid
`20:4 n-6
`g/100g lipid
`20:5 n-3
`g/100g lipid
`21:5 n-3
`g/100g lipid
`22:0
`g/100g lipid
`22:1 n11+9+7
`g/100g lipid
`22:4 n-6
`g/100g lipid
`22:5 n-3
`g/100g lipid
`22:6 n-3
`g/100g lipid
`24:1 n-9
`g/100g lipid
`
`04.01.04
`5.0
`13.9
`4.0
`0.6
`0.1
`0.5
`0.6
`11.9
`1.0
`0.5
`0.1
`1.1
`<0.1
`0.5
`<0.1
`<0.1
`<0.1
`0.2
`0.2
`13.7
`0.4
`<0.1
`0.2
`<0.1
`0.2
`8.4
`<0.1
`
`16.01.04
`5.1
`13.5
`2.8
`0.5
`0.1
`0.2
`0.5
`12.2
`1.0
`0.9
`0.1
`2.1
`<0.1
`0.5
`<0.1
`0.1
`<0.1
`0.2
`0.2
`10.1
`0.2
`<0.1
`0.2
`<0.1
`0.2
`7.5
`<0.1
`
`23.01.04
`3.2
`11.2
`1.4
`0.4
`<0.1
`0.1
`0.3
`9.7
`1.2
`1.3
`0.1
`2.5
`0.1
`0.4
`0.1
`0.1
`<0.1
`0.3
`0.3
`10.5
`0.3
`<0.1
`0.1
`<0.1
`0.3
`9.8
`<0.1
`
`24.01.04
`3.5
`12.8
`1.6
`0.5
`<0.1
`0.1
`0.4
`10.8
`1.3
`1.4
`<0.1
`2.9
`0.1
`0.4
`0.1
`0.1
`<0.1
`0.3
`0.3
`12.2
`0.3
`<0.1
`0.1
`<0.1
`0.3
`11.2
`<0.1
`
`27.01.04
`3.8
`13.3
`1.7
`0.5
`0.1
`0.2
`0.4
`11.1
`1.3
`1.4
`0.1
`3.1
`0.1
`0.4
`0.1
`0.1
`<0.1
`0.3
`0.3
`13.0
`0.3
`<0.1
`0.1
`<0.1
`0.3
`11.8
`<0.1
`
`Total saturated
`Total monoene
`Total PUFA
`Total PUFA (n-3)
`Total PUFA (n-6)
`Total fatty acids
`
`EPA/DHA
`
`g/100g lipid
`g/100g lipid
`g/100g lipid
`g/100g lipid
`g/100g lipid
`g/100g lipid
`
`19.6
`16.6
`26.9
`24.5
`1.3
`63.1
`
`19.1
`15.7
`23.4
`21.3
`1.3
`58.2
`
`14.8
`11.7
`27.2
`25.0
`1.6
`53.7
`
`16.7
`12.8
`31.0
`28.8
`1.7
`60.5
`
`17.5
`13.2
`32.8
`30.4
`1.7
`63.6
`
`1.63
`
`1.35
`
`1.07
`
`1.09
`
`1.10
`
`The EPA/DHA ratio is from 1 to 1.6, lower than in krill oil (Table 11).
`
`12
`
`AKER EXHIBIT 2003 PAGE 0016
`
`

`

`4.7 Physical characteristics
`Important properties for handling and storage of powders are flow, dust formation and bulk
`density.
`
`The flow is measured by a standardized pouring of powder over a cylindrical block with 5
`cm diameter until the cone stops growing. The height of the cone reflects the flow
`properties of the powder. If the height is greater then 5 cm the powder has very poor flow
`properties. If the height is less than 2 cm the powder has very good flow properties. The
`height of the powder cone is denoted Flow number.
`
`Table 8. Flow number in krill meal
`The values for fat is from Table 18
`
`Sample
`
`Flow number
`
`Trip 1 Comp. 1
`11.03.-20.03.05
`Kett duplicate
`May 28 - Jun 19
`Offload 22/8/05
`K.KOSHKIN
`
`3,0
`6,2
`4,8
`3,7
`2,6
`4,2
`
`Fat EU (w/acid hydr)
`g/100 g
`22,4
`26,3
`21,1
`18,1
`13,1
`18,5
`
`12
`
`14
`
`16
`
`20
`18
`Fat (g/100 g)
`
`22
`
`24
`
`26
`
`28
`
`6,5
`
`6
`
`5,5
`
`5
`
`4,5
`
`4
`
`3,5
`
`3
`
`2,5
`
`Flow number
`
`2
`
`10
`
`Figure 7. Flow number in krill meal versus fat content.
`× - Krill meal from K. Koshkin. - - - Regression line
`
`The flow number of krill meal is dependent on the fat content (Table 8 and Figure 7). Four
`of the five samples from Atlantic Navigator lie almost on a straight line when plotted
`against fat content. The krill meal with the highest fat content did not produce a cone. The
`value given is the highest heap that could be formed before it broke. The value still
`correlates well with the other samples. The meal from Konstruktor Koshkin follows the
`same relation as meal from Atlantic Navigator.
`
`The regression line for the four samples from Atlantic Navigator is
`Flow
`number
`= Fat
`.0
`(%)
`13.1
`227
`. Standard error: 0.16. Correlation coefficient: 0.99.
`×
`−
`
`The sample from Trip 1-05 (Comp.1, 28.1.-6.2.) has a much better flow (lower flow
`number) than should be expected from the fat content. The reason for this discrepancy is
`not known.
`
`13
`
`AKER EXHIBIT 2003 PAGE 0017
`
`

`

`4.8 Kett Composition Meter
`The mean values of the Kett KJT270 are compared to composite samples analysed at
`Fiskeriforskning in Table 2. The composite samples were made from an equal amount of
`day samples, so the mean values from the Kett Analyser were not weighted with the
`production volume. Compared to the analytical results, the Kett analyser is high for
`protein, low ash, and low fluorine. The results for moisture, fat and salt are in agreement.
`
`Table 9. Kett Composition Meter
`
`F-calc is fluorine calculated by
`
`Fluorine
`
`(%)
`
`=
`
`.0
`
`0334
`
`×
`
`(
`
`Ash
`
`−
`
`Salt
`
`−
`
`.0
`
`127
`
`Fat
`
`12.10.04
`
`Trip 2 05.04.05
`
`Trip 2 11.03.-20.03.05
`
`Trip 2 21.03.-07.04.05
`
`Trip 2 11.03.-07.04.05
`
`Trip 3 ATN 05-03
`
`Trip 3 06.05.2005
`
`Trip 4 May 28 - Jun 19
`
`Trip 4 Jun 29 - Jul 5
`
`Trip 5 Jul 10 - Jul 29
`
`Trip 5 Jul 30 - Aug 15
`
`Fiskeriforskning
`Kett mean
`Fiskeriforskning
`Kett
`Fiskeriforskning
`Kett mean
`Fiskeriforskning
`Kett mean
`Fiskeriforskning
`Kett mean
`Fiskeriforskning
`Kett mean
`Fiskeriforskning
`Kett
`Fiskeriforskning
`Kett mean
`Fiskeriforskning
`Kett mean
`Fiskeriforskning
`Kett mean
`Fiskeriforskning
`Kett mean
`
`6.9
`6.9
`7.1
`7.3
`7.8
`7.4
`7.1
`7.3
`
`11.0
`9.8
`11.2
`9.3
`11.5
`9.5
`11.6
`10.2
`
`)
`Ash
`
`Salt
`
`2.4
`3.4
`3.6
`3.3
`3.3
`3.3
`3.4
`
`3.7
`3.6
`3.7
`3.5
`3.7
`3.4
`3.8
`3.7
`
`F
`
`Calc
`
`0.35
`
`0.07
`
`0.10
`
`0.07
`
`0.08
`
`0.07
`
`0.08
`
`0.09
`
`0.35
`0.12
`0.06
`0.12
`0.06
`0.13
`0.05
`
`0.10
`0.06
`0.079
`0.05
`0.143
`0.04
`0.144
`0.07
`
`Fluorin
`Protein Moistur
`e
`e
`g/100 g g/100 g g/100 g g/100 g g/100 g g/100 g
`67.6
`8.0
`9.3
`12.9
`53.2
`7.0
`16.1
`16.7
`55.3
`6.0
`25.7
`10.7
`59.3
`6.5
`25.1
`9.3
`55.0
`6.7
`26.3
`10.7
`56.3
`6.8
`25.4
`10.2
`55.1
`6.6
`26.2
`10.3
`58.8
`6.9
`25.1
`9.4
`56.1
`22.1
`58.2
`25.3
`58.1
`21.4
`61.5
`21.7
`58.6
`21.1
`63.9
`20.3
`60.3
`18.1
`63.1
`16.8
`62.1
`17.1
`66.0
`16.1
`63.7
`14.7
`67.3
`13.5
`64.2
`13.4
`68.9
`12.4
`
`The Kett KJT270 is calibrated by measuring samples with known composition. These
`measurements are the basis for future analyses. This means that the uncertainty of the Kett
`analyses can never be better than, but may be as good as, the chemical methods used for
`the calibration samples. Values outside the range of the calibration samples have a greater
`uncertainty, and the calibration should be extended when samples with extreme values
`appear.
`
`The Kett KJT270 is based on the absorption of infrared radiation by protein, fat, and water.
`Ash
`otein
`Fat
`Water
`. The
`The instrument can be used to determine ash since
`100
`Pr
`≈
`−
`−
`−
`variation in salt in the krill meal samples is limited (Figure 2, Table 18) so the Kett
`KJT270 can give the same number every time and still be in agreement with the chemical
`analyses. Fluorine is correlated with salt free ash, so its determination by the instrument is
`based on the same principle as that for ash.
`
`14
`
`AKER EXHIBIT 2003 PAGE 0018
`
`

`

`Figure 8. Fat in krill meal determined on
`board Atlantic Navigator by Kett
`KJT270
`
`Figure 9. Protein in krill meal
`determined on board Atlantic Navigator
`by Kett KJT270
`
`Figure 10. Moisture in krill meal
`determined on board Atlantic Navigator
`by Kett KJT270
`
`4.8.1 Kett analyses 2005
`
`30
`
`25
`
`20
`
`15
`
`10
`
`Fat %
`
`Trip 1
`
`Trip 2
`
`Trip 3
`
`Trip 4
`
`Trip 5
`
`5
`24.01.05
`
`21.02.05
`
`21.03.05
`
`18.04.05
`
`16.05.05
`
`13.06.05
`
`11.07.05
`
`08.08.05
`
`75
`
`70
`
`65
`
`60
`
`55
`
`Protein %
`
`50
`24.01.05
`
`Trip 1
`
`Trip 2
`
`Trip 3
`
`Trip 4
`
`Trip 5
`
`21.02.05
`
`21.03.05
`
`18.04.05
`
`16.05.05
`
`13.06.05
`
`11.07.05
`
`08.08.05
`
`10
`
`9,5
`
`9
`
`8,5
`
`8
`
`7,5
`
`7
`
`6,5
`
`6
`
`5,5
`
`Moisture %
`
`5
`24.01.05
`
`Trip 1
`
`Trip 2
`
`Trip 3
`
`Trip 4
`
`Trip 5
`
`21.02.05
`
`21.03.05
`
`18.04.05
`
`16.05.05
`
`13.06.05
`
`11.07.05
`
`08.08.05
`
`The fat content of krill is high in summer, than reduces during spawning and winter. The
`fat conten