SC/66b/SP

Field and analytical protocols for the comparison of using
lethal and non-lethal techniques under the JARPNII withpreliminary application to biopsy and faecal sampling
TOSHIHIRO MOGOE1, TSUTOMU TAMURA1, HIDEYOSHI YOSHIDA2, TOSHIYA KISHIRO3, GENTA YASUNAGA1,
TAKEHARU BANDO1, TOURU KITAMURA4, NAOHISA KANDA4, KOICHIRO NAKANO5, HIROSHI KATSUMATA5,YOSHIHIRO HANDA5 AND HIDEHIRO KATO6
1

Institute of cetacean research, 4-5 toyomi-cho, chuo-ku, Tokyo 104-0055, japan
National Research Institute of Far Seas Fisheries, 2-12-4 Fukuura, Kanazawa, Yokohama, Kanagawa 2368648, Japan
3
Headquarters of Japan Fisheries Research and Education Agency, 15F Queen's Tower B, 2-3-3 Minato Mirai,
Nishi-ku, Yokohama, Kanagawa, 220-6115, Japan
4
JAPAN NUS Co., Ltd., 7-5-25 Nishi-Shinjuku, Shinjuku-Ku, Tokyo 160-0023, Japan
5
Seibutsu Giken Co., Ltd., 3068 Sakai, Atsugi-shi, Kanagawa 243-0022, Japan
6
Tokyo University of Marine Science and Technology, 5-7 Konan 4, Minatoku, Tokyo 108-8477, Japan
Contact e-mail: mogoe@cetacean.jp
2

ABSTRACTIn response to the report of the Expert Panel for the final review of the western North Pacific Japanese Special
Permit programme (JARPNII), we provide the field and analytical protocols for the comparison of using lethal
and non-lethal techniques. The study is conducted for three years in the coastal water off Sanriku and Kushiro, andoffshore. Primary questions are (1) whether a tissue and other samples can be obtained by a non-lethal method;(2) whether enough samples for statistical analysis can be obtained by the non-lethal method; (3) whether the
sample obtained by the non-lethal method can produce scientific information compared to that produced by a lethal
sampling method; and (4) whether the cost for obtaining the sample/producing scientific information is reasonable.
As a result of preliminarily applying the field and analytical protocols to the data obtained in 2014 and 2015, we
provisionally concluded that sampling efficiency of faeces was very low, and also that the estimation based on
DNA analysis are unreliable at this stage because prey species different from stomach contents were identified
from DNA analyses of large intestine. As for biopsy sampling, the samples could be obtained from swimming
animals, though sampling efficiency will be different by species. We will evaluate the feasibility and practicabilityof non-lethal means by applying the same approach, using data in the comparative study including those obtained
this year.
KEYWORDS: SCIENTIFIC PERMITS; COMMON MINKE WHALE; BRYDE’S WHALE; SEI WHALE

INTRODUCTION
The decision to finalize the JARPNII program was made taking account of the International Court of Justice (ICJ)
Judgment in the case concerning Whaling in the Antarctic (Australia v. Japan: New Zealand intervening). The ICJ
Judgment on 31 March 2014 states that “It is to be expected that Japan will take account of the reasoning andconclusions contained in this Judgment as it evaluates the possibility of granting any future permits under Article
VIII, paragraph 1, of the Convention” (paragraph 246). Therefore, while JARPNII was not included within thesubject matter of the case, the Government of Japan reviewed the design of the ongoing JARPN II under its
decision described in the Statement by Ministry for Agriculture, Forestry and Fisheries on 18 April 2014.
The purpose of the changes in sample sizes includes conducting comparative research between lethal and nonlethal methods. With regard to implications of the adjustment for meeting the objectives of the programme, thecomparative research is designed to verify a hypothesis that lethal methods could be replaced by non-lethal
methods. In other words, if non-lethal methods adopted since 2014 produce equivalently useful data as the lethal
method, there will be no hindrances to meeting the original objectives of the program. While the proponents willsubmit the provisional results of the comparative research to the 2016 IWC SC, the results of the verification will
be reported after the three years (2014-2016) research period.

1

The review panel for JARPNII recommended that the proponents provide the field and analytical protocols for thecomparison of using lethal and non-lethal techniques for each key parameter taking into account the advice
provided in 2009 (IWC, 2016). Following the preliminary response by the proponents that they would provide theprotocols at the SC 66b meeting (Tamura et al, 2016a), we provide the field and analytical protocols to evaluate
the feasibility and practicability of non-lethal means, especially biopsy and faecal sampling under the prioritizedobjective. We also provide results obtained to date, which is an update of the preliminary results reported to theSC 66a meeting, and apply the protocol to the data available.

FIELD AND ANALYTICAL PROTOCOLS
1. Procedure for sampling
Biopsy sampling
The equipment for biopsy skin and/or blubber tissue were a crossbow or Larsen gun system (Larsen, 1998). The
open sight of Larsen gun system was replaced with an electronic aiming device (red-dot-sight), which allows fasteraiming and thus faster shooting. The biopsy darts (4 inches) consists of a carbon fibre shaft, which is high-pressuremoulded to a polyethylene float that also functions as a stop to limit penetration into the tissue. In the float end of
the dart, a threaded insert is used for attaching the screw-on biopsy-sampling tip. The biopsy tip is a stainlesscylinder with a 9mm outer diameter, an internal diameter of 7mm and three internal barbs for sample retention. To
avoid sampling failure or to improve the sensitivity of analysis, re-sampling from the identical individuals are
conducted. These samples should be independently collected and not divided after sampling to increase the sample
size. Information on time taken, sea state, and swell was recorded to enable a plausible measure of effort to be
developed. The researcher should remove the specimen from the biopsy-sampling tip with a sterile needle orsingle-use tweezers. And place it in a 99% ethanol filled tube or in a small zip plastic bag (−20 ºC). Each of thespecimens is separately labelled with sample names (e.g. J16YS1M001).

Faecal sampling
Observation of excretion from the identified whales and sampling of excreted faeces were conducted for common
minke, sei and Bryde’s whales. The observed time is defined as the duration of approaching to the whale within
0.2 n.mile (confirmation of whale species) to end of chasing or observation. If an observer found faeces near thesurface of the sea water, the faeces were sampled by circle net with 100μm mesh size. The sampled faeces werestored using polyethylene bottles at −20°C.

Blubber samples
In order to answer Q3 below, it is useful to compare data from the same animal which can be obtained only lethally(e.g. stomach contents) to that which could be obtained non-lethally (e.g. blubber tissue sample). For that purpose,skin and blubber tissue samples from lethal techniques are stored at −20°C.

Contents of large intestines
In the same vein, for an experiment to determine the practicability of contents of large intestines contents of large
intestines are collected from the lethally sampled whales and stored at −20°C.

2. Preparation of data from lethal and non-lethal samples
Tissue samples from skin and blubber
Stable isotope and fatty acid analysis on the skin and blubber samples are performed to evaluate their comparability
to stomach content data. The stable isotope analysis is conducted in collaboration with the Japan Chemical
Analysis Centre, Chiba Prefecture, Japan.

Faecal samples and contents of large intestines
DNA analyses on faecal samples and contents of large intestines are performed to evaluate their comparability tostomach content data. Total genomic DNA of each individual is extracted using the standard phenol/chloroform

2

extractions protocol of the GENTRA PUREGENE DNA extraction kit (QIAGEN) following the company’s
manual. Extracted DNAs are stored in the TE buffer. After PCR amplification, the products are analysed using an
Illumina MiSeq (Next-generation DNA sequencers) to identify the prey species. These results are compared tothose observed from stomach content on an individual base.

Stomach content data
The data is obtained from the analysis of stomach contents. Details of the analysis are given in Tamura et al,(2016b).

3. Comparison of lethal and non-lethal means/samples (common for biopsy and faecal sampling)
Proponents specified the following four questions to be answered in order to evaluate the feasibility andpracticability of non-lethal means, especially biopsy and faecal sampling under the prioritized objective (Mogoe
et al, 2015);(1) whether a tissue and other samples can be obtained by a non-lethal method (e.g. biopsy sampling, faeces
collection);(2) whether enough number of samples for statistical analysis can be obtained by the non-lethal method;(3) whether the sample obtained by the non-lethal method can produce scientific information comparable to that
produced by a lethal sampling method; and(4) whether the cost for obtaining the sample/producing scientific information is reasonable.
Questions 1 and 2 above are technical, Question 3 is analytical, and Question 4 is a logistical evaluation.

Systematic application of the questions
The fundamental question as to whether research objectives are achievable non-lethally can be answered throughthese questions. A flow chart (see Figure 1) articulates how these four questions are applied to the biopsy andfaecal sampling/samples, and specifies the condition where we can conclude biopsy and/or faecal sampling canreplace lethal take. The systematic application of the questions forms a basis to objectively discuss the feasibility
and practicability of non-lethal means, particularly from a perspective of whether research objectives areachievable through non-lethal means. The objective basis of discussion precludes repetitive discussion as to thefeasibility and practicability of non-lethal means, which has taken place to date.

Criteria to answer each of the questions
For Question 1, the criteria is simple. If at least one sample could be taken during the research period, the answer
is yes and otherwise the answer is no.
For Question 2, sampling efficiency needs to be compared between lethal and non-lethal means. According to thepast studies, biopsy sampling efficiency was examined by the number of samples obtained per shot (Isoda et al,2016). In this study, we regard sampling efficiency as a combination of effort to obtain samples and success rate.
Effort and success rate are defined as follows:

Effort: Sum of time from 'confirmation of whale species' to 'obtain sample' for whales successfully sampled+ Sum of time from 'confirmation of whale species' to ‘lose whale’ for those targeted but not sampled
Success Rate:
Success Rate 1 (# of sampled whales/＃of targeted whales)
Success Rate 2 (# of samples/ # of shots for each whale)

Question 2 can be answered by taking account of all these results.

3For Question 3, whether prioritized research objectives, i.e. prey consumption, prey preference, and ecosystemmodelling, can be achieved with data from non-lethal means needs to be examined. Under these objectives, it isrequired that species composition in diet is quantified accurately and that information is available as model inputs.
This aspect will be examined with data obtained from non-lethal means.
For Question 4, reasonableness can be evaluated by dividing the overall cost for research by the number of samples
obtained.

APPLICATION OF THE PROTOCOL TO THE DATA OBTAINED
Effort for non-lethal research activity
Non-lethal survey activities were carried out in three different areas within the 2014 and 2015 JARPNII. Table 1
shows a searching ‘on effort’ (time) with the data obtained by the each research activity. The Sighting andSampling Vessels (SSV) of offshore components were Yushin Maru type (Bando et al, 2016). Different survey
platforms (research vessels) were used for the Offshore and Coastal components. Smaller vessels were used in the
coastal survey (Mogoe et al, 2016; Kishiro et al, 2016). The Sighting Vessels (SVs) used were Yushin Maru typein the sighting survey of offshore components. Total searching time of non-lethal and lethal were 1,238.4 and
2,134.1 hours, respectively. Percentages of non-lethal effort were from 8.4 to 61.8%, except for SV data.

Sampling efficiency of biopsy and faecal sampling (Q1 and Q2)
Data obtained from biopsy and faecal sampling in 2014 and 2015 were used for preliminary evaluation of sampling
efficiency.
Of a total of 157 biopsy sampling attempts, we collected 114 samples (38 samples from sei, 68 from Bryde’s, and8 from common minke whales). Table 2 summarises the success rates of biopsy sampling. Success rate 1 for thesei, Bryde’s and common minke whales were 47.1%, 71.6% and 38.1%, respectively. These results indicate thatbiopsy samples are obtainable from the species, that is, the answer to the Question 1 is ‘YES’, for all species. The
results, however, suggests that sampling efficiency will be different among the species. Biopsy sampling effort(time) was compared with the effort required for obtaining lethal samples in each species. The average samplingtime of sei whales was 55.7 minutes for biopsy sampling, while it was 31.4 minutes for lethal sampling; for Bryde’s
whales, the average sampling time was 26.8 minutes for biopsy sampling, while it was 25.6 minutes for lethal
sampling; for common minke whales off Kushiro, the average sampling time was 149.6 minutes, while it was 72.1
minutes for the lethal sampling. Sampling efficiency of biopsy sampling is thought to be higher for Bryde’s whales
than that for the other two species. We will evaluate efficiency of biopsy sampling by species, by adding dataobtained from the 2016 surveys, to answer the Question 2.
Regarding the faecal sampling, proponents conducted a total of 1808 experiments (1,179 for sei, 393 for Bryde’s
and 236 for common minke whales), for 377.2 hours (Table 3). Throughout the experiments, excretion wasobserved for 38 individuals (30 for sei, 6 for Bryde’s, and 2 for common minke whales). Of these, faeces was
obtained successfully only from 5 sei whales. For Bryde’s and common minke whales, faeces sampling was failed,
due to sink or spread of faeces before sampling. Our results indicate that, at the present, the answer to the Question
1 is ‘YES’, only for sei whales. Table 3 shows that encounter rate with faeces at the sea is extremely low (2.1% of
all experiments). Sampling efficiency of faecal sampling from swimming animals will not be so high, especiallyfor Bryde’s and common minke whales. The efficiency will be evaluated further, by adding the 2016 survey data.

Comparison of data between faecal samples and stomach contents (Q3)
The studies on faecal steroid metabolites published for free-living whales are limited by the difficulty in obtaining
samples except for North Atlantic right whales (Gillett et al, 2008). Zooplankton in faecal samples should be
identified using by a dissecting microscope to determine species composition, quantity and developmental stages(Swaim et al, 2009). However, faecal contents of sei and Bryde’s and common minke whales showed that thecontents derived from zooplankton (such as copepods) tend to float while the faeces from whales feeding fish tend
to sink (Mogoe et al., 2015).
Preliminary findings from the DNA analyses of contents of intestine and faeces using next-generation sequencing(NGS) technologies (Table 4) are as follows:
1. The prey species compositions identified in the contents of large intestine were quite different from thosein the stomach contents.

4

2. Although this method provides some prey information even from a whale without stomach contents, insome cases, no prey species were identified in the contents of large intestine of the individuals
notwithstanding the full stomach contents. In addition, the prey identification rate differed among thewhale species.
3. Prey of the prey species were also detected (e.g., Copepoda: Acartia clausii in the individual 14NPCSM019 is known as a major prey of sand lance and not as a prey of common minke whales). Suchcontamination of prey of prey species causes a critical problem in feeding ecology study of whales.
4. The results of prey ID from the contents of upper part and middle part in the small intestine were similar
to contents of the stomach rather than that of the large intestine.
5. The prey ID could not be obtained from faecal samples of sei whales by using NGS.
The next step to be conducted especially for points 1 and 2 above is to examine whether these results were due toeither biological, technical or both reasons. One of the technical reasons which we encountered in this study wasthat almost all of the PCR products predominantly contained the fragments of the whale sequences, causing low
prey species identification rate in the samples. Development of a method which avoids amplifying the whale
sequences (e.g., Shehzad et al, 2012) is now under way, so a better resolution will be obtained in near future.
Likewise, a solution should be found to separate amplification of prey of the prey species. This preliminary study
clearly indicates that the genetic prey ID only in the contents of large intestine (and faeces) is insufficient tounderstand feeding habits of the whales at this stage.
With regard to the comparison between stable isotopic data and stomach contents data, according to Icelandic
research whaling in the North Atlantic, results from the stable isotope analysis showed a considerably lower trophic
level in food consumption compared to the trophic levels indicated by the stomach content analysis (Víkingsson
et al, 2013). They concluded that depending solely on stable isotope results, in general, may not be able to give an
accurate profile of diet in the case of a highly generalist predator such as common minke whales (Ólafsdóttir et al,2013).
Currently, stable isotope analyses on skin/blubber samples are being conducted and proponents will provide
results of the comparison between stable isotopic data and stomach contents data in the North Pacific.

CONCLUSION
Proponents developed the field and analytical protocols specifically for the comparative study, but they are also
universally applicable to evaluate the feasibility and practicability of non-lethal means. Systematic application of
the protocols are an efficient and constructive way because, even though the feasibility and practicability of nonlethal means have been repeatedly discussed, the conclusion was often obscure due to a lack of an objective
evaluation scheme.
A few faeces of sei whales were obtained, whereas no faeces of Bryde's and common minke whales were obtained.
Furthermore, encounter rates of faeces at sea in the three species were extremely low. Compositions of prey species
based on our DNA analysis of large intestine contents of whales were quite different from those based on
observation of stomach contents. And also, we could not identify prey species from faeces of three sei whales.
Therefore, sampling efficiency of faeces was very low, and also the estimation based on DNA analysis areunreliable at this stage.
As for biopsy sampling, we could obtain the samples from all the three species by using Larsen gun. The results
suggest that the answer to the Question 1 is ‘YES’ to date. Our results also suggest that sampling efficiency ofbiopsy sampling will be different among the species. The sampling for Bryde’s whales is thought to be more
efficient than that for the other two species. We will collect further data, because our data are insufficient. At thesame time, we will start to investigate estimation of prey composition and trophic level using skin sample of
JARPNII. Proponents will evaluate biopsy sampling by applying the same approach, using data obtained in thecomparative study, in addition to the data collected in 2016 surveys.

ACKNOWLEDGEMENTS
We would like to thank all captains, crews and researchers, who were involved in the JARPN II surveys. Our
sincere thanks to Messrs. Naohito Okazoe and Hiroyuki Morita, the Fisheries Agency of Japan for their valuablesuggestions and useful comments on this paper.

5

REFERENCES
Bando, T., Isoda, T., Nakai, K., Sato, H., Okitsu Y., Tsunekawa, M., Fujii, K., Yamadaki, M., Honma, H andEguchi, K. 2016. Cruise Report of the Second Phase of the Japanese Whale Research Program under Special
Permit in the Western North Pacific (JARPN II) in 2015 (part I) - Offshore component - . Paper SC/66b/SP
presented to the 66b IWC Scientific Committee, May 2016 (unpublished).
Gillett, R.M., White, B.N. and Rolland, R.M. 2008. Quantification and genetic profiling of DNA isolated fromfree-floating feces of the North Atlantic right whale (Eubalaena glacialis). Mar. Mamm. Sci. 24: 341–55.
International Whaling Commission. 2016. Report of the Expert Panel of the final review on the western North
Pacific Japanese Special Permit programme (JARPN II). Paper SC/66b/Rep06 presented to the IWC
Scientific Committee, June 2016 (unpublished). 96pp.
Isoda, T., Kawabe, S., Ohkoshi, C., Mogoe, T. and Matsuoka, K. 2016. Results of the NEWREP-A dedicatedsighting survey in Area IV during the 2015/16 austral summer season. Paper SC/66B/IA presented to the
IWC Scientific Committee, June 2016 (unpublished). 25pp.
Kishiro, T., Yoshida, H., Ito, N., Mogoe, T., Nakamura, G., Maeda, H., Miyakawa, N., Hirose, A., Ota, M., Kato,
K., Hayashi, R., Yoshii, K., Kim, Y., Miyashita, T., Kumagai, S., Sato, H., Kimura, Y., Hiruda, H., Katsumata,T., Nakai, K., and Kato, H. 2016. Cruise report of the second phase of the Japanese Whale Research Program
under Special Permit in the western North Pacific (JARPN II) in 2015 (Part III) – Coastal component off
Kushiro. Paper SC/66b/SP paper presented to the IWC Scientific Committee, June 2016 (unpublished). 12pp.
Larsen, F. 1998. Development of a biopsy system primarily for use on large cetaceans. Paper SC/50/O15
presented to the IWC Scientific Committee, May 1998 (unpublished). 8pp.
Mogoe, T., Tamura, T., Yoshida, H., Kishiro, T., Yasunaga, G., Bando, T., Konishi, K., Nakai, K., Kanda, N.,
Kitamura, T., Nakano, K., Katsumata, H., Handa, Y. and Kato, H. 2015. Preliminary report of efficiency andpracticability of biopsy sampling, faecal sampling and prey species identification from genetic analyses in
2014, and workplan for non-lethal research in JARPN II. Paper SC/66a/SP11 presented to the IWC
Scientific Committee, May 2015 (unpublished). 11pp.
Ólafsdóttir, D., Víkingsson, G.A., Elvarsson, B. and Auðunsson, G.A. 2013. Analyses on stable carbon andnitrogen isotope ratios in soft tissues of common minke whale (Balaenoptera acutorostrata) in Icelandic
waters and its prey. SC/F13/SP3 to the IWC Scientific Committee. 13pp.
Shehzad, W., McCarthy, T.M., Pompanon, F., Purevjav, L., Coissac, E., Riaz, T. and Taberlet, P. 2012. Prey
preference of snow leopard (Panthera uncia) in South Gobi, Mongolia. PLoS One 7(2):e32104. doi:
10.1371/journal.pone.0032104.
Swaim, Z.T., Westgate, A.J., Koopman, H.N., Rolland, R.M. and Kraus, S.D. 2009. Metabolism of ingested
lipids by North Atlantic right whales. Endang. Species Res. 6:259–71.
Tamura, T., Kishiro, T., Bando, T., Yasunaga, G., Murase, H., Maeda, H., Kitakado, T. and Pastene, L.A. 2016a.
Response to the ‘Report of the Expert Panel of the final review on the western North Pacific JapaneseSpecial Permit program (JARPNII)’. Paper SC/66b/SP01 presented to the IWC Scientific Committee, June
2016 (unpublished). 11pp.
Tamura, T., Konishi, K. and Isoda, T. 2016b. Updated estimation of prey consumption by common minke,
Bryde’s and sei whales in the western North Pacific. Paper SC/F16/JR15 presented to the Expert Panel ofthe final review on the western North Pacific Japanese Special Permit programme (JARPN II), Tokyo,
February 2016 (unpublished). 58pp.
Víkingsson, G.A., Elvarsson, B., Chosson, V., Ólafsdóttir, D. and Galan, A. 2013. Recent changes in the dietcomposition of common minke whales (Balaenoptera acutorostrata) in Icelandic waters. – Consequence ofclimate change? SC/F13/SP2 to the IWC Scientific Committee. 31pp.

6

Figure 1. Systematic application of the four questions to evaluate non-lethal means. It should be noted that whenat least one of the arrows reaches the box “lethal take is necessary to achieve research objectives,” theProponents conclude that lethal take is necessary for the research program.

Table 1. Searching effort time (hours) of each survey.
Offshore SSVs

Offshore SVs

Coastal Sanriku

Coastal Kushiro2014 JARPNII
Lethal effort
Non-lethal effort
Total effort
Rate (%) of non-lethal effort

193.3
89.6
282.9
31.7

—
262.4
262.4
100

510.3
60.8
571.1
10.6

250.9
58.6
309.5
18.9

2015 JARPNII
Lethal effort
Non-lethal effort
Total effort
Rate (%) of non-lethal effort

61.5
99.5
161
61.8

—
547.5
547.5
100

596.4
54.6
650.9
8.4

521.7
65.4
587.1
11.1

7

Table 2. Success rates and effort required for one sample under biopsy sampling in the 2014 and 2015 surveys.
Species2014 JARPN II
Sei
Bryde's
C. minke (Offshore)
C. minke (Sanriku)
C. minke (Kushiro)
2015 JARPN II
Sei
Bryde's
C. minke (Offshore)
C. minke (Sanriku)
C. minke (Kushiro)
Total
Sei
Bryde's
C. minke (Offshore)
C. minke (Coastal)

Ship Number of Targeted Number of Number of Number of Number of
type
Exp.
whalesshootshits
samples* sampledwhales(A)(B)

Effort(min)

Success
rate 1

Success
rate 2

(C)

(D)

(E)

(D)/(A)(C)/(B)

Effortrequiredfor onesample(min)(E)/(D)

SSVs
SVs
SSVs
SVs
SSVs
SVs
SSVs
SSVs

33
0
37
0
0
0
0
9

42
—
39
—
—
—
—
9

63
—
67
—
—
—
—
14

21
—
31
—
—
—
—
5

16
—
25
—
—
—
—
5

16
—
25
—
—
—
—
5

1,275
—
789
—
—
—
—
458

0.381
—
0.641
—
—
—
—
0.556

0.254
—
0.373
—
—
—
—
0.357

80
—
32
—
—
—
—
92

SSVs
SVs
SSVs
SVs
SSVs
SVs
SSVs
SSVs

25
0
41
0
2
2
1
7

26
—
42
—
2
2
1
7

44
—
90
—
4
2
1
4

22
—
46
—
2
2
0
4

22
—
43
—
2
1
0
0

16
—
33
—
2
1
0
0

507
—
763
—
52
20
54
236

0.615
—
0.786
—
1.000
0.500
0.000
0.000

0.364
—
0.367
—
0.500
0.500
0.000
0.000

32
—
23
—
26
20
—
—

58
78
4
17

68
81
4
17

107
157
6
19

43
77
4
9

38
68
3
5

32
58
3
5

1,782
1,552
72
748

0.471
0.716
0.750
0.294

0.299
0.369
0.500
0.263

56
27
24
150

Table 3. The results of faecal sampling in the 2014 and 2015 surveys.
Species2014 JARPA II
Sei
Bryde's
C. minke (Offshore)
C. minke (Sanriku)
C. minke (Kushiro)
2015 JARPA II
Sei
Bryde's
C. minke (Offshore)
C. minke (Sanriku)
C. minke (Kushiro)
Total
Sei
Bryde's
C. minke (Offshore)
C. minke (Coastal)

Ship type

Number ofexperiments(school)

Number ofexperiments(individuals)

SSVs
SVs
SSVs
SVs
SSVs
SVs
SSVs
SSVs

192
134
94
30
2
2
49
89

346
333
116
42
2
2
49
89

75.1
5.9
25.4
12.7
0.1
0.2
44.8
60.6

11
10
1
2
0
0
0
1

3
0
0
0
0
0
0
0

SSVs
SVs
SSVs
SVs
SSVs
SVs
SSVs
SSVs

193
133
113
70
2
0
33
59

259
241
147
88
2
0
33
59

51.6
7.7
27.4
2.4
0.9
0.0
31.0
31.4

6
3
2
1
0
0
0
1

2
0
0
0
0
0
0
0

652
307
6
230

1,179
393
6
230

140.3
67.9
1.2
167.8

30
6
0
2

5
0
0
0

8

Observation effort Observation of(hours)excretion(number)

Faecal sampling(number)Table 4. Results of detected prey species in enteral content using next generation DNA sequencers.
Species

ID numberPrey species observed by Prey species estimated by Prey species estimated by Prey species estimated bystomach contents
NGS – Upper part of small NGS – Middle part of
NGS – Large intestine
intestine
small intestineSei

14NPSE001

Mackerels (90%) andMackerels and JapaneseJapanese anchovy (10%) anchovy
Copepods (99%) and krill Krill(1%)
Mackerels
Mackerels and PacificsauryJapanese sardine (50%), Japanese sardine andJapanese anchovy (40%) Japanese anchovy
and Mackerels (10%)
Copepods (80%) and
Pacific saury
Pacific saury (20%)
Copepods
Pacific saury
Copepods
No identified
Mackerels
Mackerels and Pacificsaury

Mackerels and Japanese
anchovy
No identified

No identified

Pacific saury

No identified

No identified

No identified

Pacific saury

Krill

Pacific saury
No identified
Mackerels

Pacific saury
No identified
No identified

14NPB005
14NPB006

Japanese anchovy
Mackerels

Japanese anchovy
Japanese anchovy

Japanese anchovy
No identified

14NPB009

Japanese anchovy (90%), Japanese anchovy
Japanese sardine (8%) andMackerels (2%)
Japanese anchovy
Japanese anchovy
Japanese anchovy
Japanese anchovy
Japanese anchovy (99%) Japanese anchovy
and mackerels (1%)

Japanese anchovy

No identified
Light fish (Maurolicus
muelleri)Japanese anchovy,
Japanese sardine and krill

No identified
Japanese anchovy
Japanese anchovy

No identified
Krill
No identified

Sand lance
Sand lance
Sand lance
Japanese sardine
Japanese sardine
Japanese sardine
Walleye pollock andJapanese sardine
Japanese sardine
Walleye pollock andJapanese common squid

—
—
—
—
—
—
—

—
—
—
—
—
—
—

Sand lance
Copepoda (Acartia clausii)
No identified
No identified
No identified
Japanese sardine
No identified

—
—

—
—

Japanese anchovy
Japanese common squidand krill

14NPSE006
14NPSE018
14NPSE044
14NPSE048
14NPSE052
14NPSE067
14NPSE070

Bryde’s

14NPB010
14NPB016
14NPB019

C.minke

14NPCS-M013
14NPCS-M019
14NPCS-M021
14NPCK-M013
14NPCK-M015
14NPCK-M016
14NPCK-M017
14NPCK-M019
14NPCK-M027

Krill

NGS: next-generation sequencing.

Table 5. Results of detected prey species in faeces using next generation DNA sequencers.
Species

ID Number

Results

Sei

140527SEI

Oithona similisSei

140528SEI

Oithona similisSei

150529SEI

Euphausiacea, Calanoida

9