SC/D06/J32

OVERVIEW OF OTHER STUDIES ON LARGE WHALESBASED ON JARPA SAMPLES
Luis A. Pastene, Hajime Ishikawa, Mutsuo Goto and Shigetoshi Nishiwaki
Institute of Cetacean Research, Toyomi-cho 4-5, Chuo-ku, Tokyo 104-0055, Japan

ABSTRACTJARPA collected systematically a comprehensive data/sample set that allowed the conduction ofstudies related with the four objectives of the program. Samples collected by both lethal and non-lethal
components of JARPA have been also used in several studies with objectives different from the four
objectives of the program. In this paper an overview of these studies are conducted. They havecontributed to the whale knowledge mainly in three areas: reproductive physiology of Antarctic minke
whale, phylogeny of large whales and distribution and movement of baleen whales.
INTRODUCTION
Samples and data obtained systematically during the Japanese Whale Research Program under Special
Permit in the Antarctic (JARPA) have been collected mainly with the purpose to conduct studies
related with the four objectives of the JARPA (SC/D06/J1). Samples and data collected by both lethal
and non-lethal components of the JARPA have been also used in several studies with objectives
different from the four objectives of the program.
These ‘additional’ studies have been reported annually to the Scientific Committee meeting throughthe annual report of the Institute of Cetacean Research (ICR) research activities (RAICR). Details of
the studies can be found in the original papers listed in the web page
http://www.icrwhale.org/JARPAReview1.htm, and in the references listed below.
The objective of this paper is to conduct an overview of the studies conducted using JARPA
sample/data, which are not related to the main objectives of the program. The overview is focused
mainly to three areas of research contribution: reproductive physiology of Antarctic minke whale,
phylogeny of large whales and movement and distribution of baleen whales. It should be noted that insome of these studies JARPA sample/data have been a part of large data sets used.
REPRODUCTIVE PHYSIOLOGY OF ANTARCTIC MINKE WHALES
These studies have been conducted in cooperation between ICR and Obihiro University ofAgriculture and Veterinary Medicine in different steps since 1994. The first step in the studyof reproductive physiology in the Antarctic minke whales involved the following topics: i)studies on relationship between appearance of preantral follicles in the fetal ovary and hormone
concentrations in the fetal heart, umbilical cord, and maternal blood, ii) studies on hormonal status inblood, ovary, and pituitary gland in immature and mature whales, and iii) studies on differentiation,function and morphology in testis. The second step involved the following topics: i) studies on in vitro
maturation, fertilization by micro-manipulation, and embryonic development of frozen-thawed or
vitrified immature oocytes, ii) studies on sperm analyses and freezing, iii) basic studies on somatic cellcloning, iv) studies on function and morphology of placenta in pregnant whales, and v) studies on milk
constituents analyses.
In the third step, the following topics were addressed: i) developmental capacity of vitrified oocytes
followed by in vitro maturation, and parthenogenetic activation or in vitro fertilization by
intracytoplasmic sperm injection, ii) the ability of whale haploid spermatogenic cells for inducing
calcium oscillations and activating oocytes, iii) contribution of spaermatozoal centrosomes tomicrotubule-organizing center in oocytes, iv) histological studies on the placenta and uterus in, v)

1studies on composition analysis and utilization as functional materials of sphingoid base lipids fromskin.
The research team successfully cryopreserved spermatozoa of Antarctic minke whales in 1994.
Relationship among physiological status of whales, serum steroid hormones and morphology offollicular oocytes and spermatozoa was also studied (Fukui et al. 1995, 1996, 2004, Iga et al. 1996,
Mogoe et al. 1998a, 1998b, 2000). After consideration on in vitro maturing (IVM) of follicular oocytesof Antarctic minke whales, they succeeded in vitro fertilization (IVF) of follicular oocytes of Antarctic
minke whales in 1996. Factors affecting each step of artificial fertilization were investigated andtechnique on cryopreservation of spermatozoa, IVM, IVF and development of oocytes have been
improved (Fukui et al. 1997a, 1997b, Asada et al. 2000, 2001a, Fujihira et al. 2004, 2006, Amemiya et
al. 2004, Fukui et al. 2004, Iwayama et al. 2004). The research team recently succeeded
intracytoplasmic sperm injection (ICSI) and interspecies somatic cell nuclear transfer (SCNT) of
Antarctic minke whales (Asada et al. 2001b, Ikumi et al. 2004) and basic study to produce cloned
whale embryos is also ongoing (Kobayashi et al. 2006).
The research team also proceeds with other reproductive studies in the Antarctic minke whale, suchas plasma and pituitary concentration of gonadotropins (FSH and LH) (Suzuki et al. 2001), chemical
characterization of the oligosaccharides in milk (Urashima et al. 2002), morphology and developmentof ovary (Tetsuka et al. 2004), relationship between preantral follicles in the fetal ovary and hormone
concentrations in fetal and maternal blood (Muranishi et al. 2004).
PHYLOGENY OF LARGE WHALESJARPA collected a number of biopsy samples from large whale species as follow: humpback whale:
342, right whale: 36, blue whale: 22; fin whale: 28 and sei whale: 1. At the laboratory all these samples
were examined genetically to obtain mitochondrial DNA (mtDNA) control region sequences and someof them (humpback and blue whales) were examined with a set of microsatellites. Sex determination
has been possible using a method developed at ICR (Abe et al. 2001).
Genetic data collected from the humpback whale were used in a genetic study on stock structure in
the feeding ground (SC/D06/J31); mtDNA sequences collected from the fin whale were used in a
genetic study focused to understand the phylogenetic relationships among fin whales from different
ocean basins (Goto et al., 2003), which was discussed during the 2003 Scientific Committee meeting.
DNA data obtained from biopsy samples in these baleen whales as well in minke whales sampledduring JARPA were used in several genetic studies focused to understand the phylogenetic
relationships among large whales as follow:
On blue whales, two collaborative genetic researches were conducted. The first was a collaboration
between ICR and the Southwest Fisheries Science Center, USA. The main purpose of the study was tostudy the genetic differentiation between pygmy and true blue whales in the Southern Hemisphere aswell among populations of the pygmy blue whales, using two genetic markers, mtDNA control region
sequences and microsatellites (LeDuc et al., in press). The second collaborative research is being
conducted between ICR and the University of California at Davis, USA. The objective of the study isthe delineation of biological stocks of blue whale worldwide using nuclear genetic markers (introns). In
both projects DNA data of blue whales from the Antarctic were from JARPA.
Regarding right whales a collaborative genetic research between the American Museum of Natural
History and several institutions included ICR was conducted to study taxonomy and phylogenetic
relationships of this species worldwide (Rosenbaum et al., 2000). MtDNA sequences of the southern
right whale used in this study were from JARPA.
DNA data of several baleen whale species from JARPA were used in a genetic study focused tounderstand positive selection in the major histocompatibility complex (MHC) variation of the cetacean
species (Hayashi et al., 2003). MHC is a large multigene coding glycoprotein, which plays a key role in
the initiation of immune responses in vertebrates.
DNA data of several baleen whale species from JARPA were used in a genetic study focused todetermine the coding region of the SRY (sex determining region on the Y chromosome) gene and its

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predicted promoter region in 20 species of cetacean. The study was also focused to construct a genetree using the SRY sequence and its predicted promoter region to determine whether the cetacean
phylogeny could be explained from the standpoint of a strictly paternal mode of inheritance (Nishida etal., 2003).
DNA data of several baleen whale species from JARPA were used to investigate the phylogeny andevolution of baleen whales based on the complete mitochondrial genome sequence (Sasaki et al., 2005)
as well short interspersed repetitive elements (SINE) (Nikaido et al., 2006).
MOVEMENT AND DISTRIBUTION OF BALEEN WHALESThe ICR has constructed a photo-ID catalog of large baleen whale species based on the mostinformative pictures taken during the JARPA surveys. The ICR photo-ID catalog involves thefollowing number of pictures: humpback whale: 502, blue whale: 153, right whale: 243. All
photographs of humpback whales have been submitted to the Centralized Antarctic Humpback Whale
Catalogue supported by the IWC and managed by the College of the Atlantic. Photo-ID data have beenused in several studies on distribution and movement of large baleen whales as follow:
Regarding right whales, analyses on photo-ID pictures are conducted in collaboration between ICR
and the Western Australian Museum, Australia. A study demonstrated movement of one animal
between Adelaide, Australia and Antarctic Area IV (Bannister et al., 1999).
Regarding humpback whale, analyses on photo-ID pictures were conducted with the purpose toinvestigate movement and distribution of this species in the Antarctic feeding grounds as well between
the Antarctic and low latitude areas. Rock et al. (in press) demonstrated movement of three individuals
between Eastern Australia and Antarctic Area V. Also one animal was identified in different years in asimilar longitudinal sector of Area VI.
OTHER
Sighting and distribution
Matsuoka et al. (1996) reported a sighting of a large school of the pygmy right whale in thesoutheastern Indian Ocean. The sighting was made by JARPA vessels during transit to the Antarctic
research area in December 1992. Kimura et al. (2005) used satellite derived ocean parameters and thewhale sighting location data collected under the JARPA to elucidate prevalent ocean condition where
whale sighting was undertaken, and to understand its influence on the distribution of the sei whales.
Lipids composition
Several studies were focused to study the fatty acid composition of blubber oils of the Antarctic as wellof the dwarf minke whales (Itoh, 1994; Itoh et al., 1990; 1993). A study focused specifically tounderstand the site and sexual differences of muscle lipids in the Antarctic minke whale (Itoh et al.,1998).
Contribution of biological materials to museums
JARPA has collected biological samples mainly for scientific research purposes. Some biologicalmaterials have been also collected and provided to museums, aquarium and university through Japanfor exhibition purposes. JARPA biological materials (especially Antarctic minke whale skeletons) havebeen provided to at least to 19 museums, aquariums or universities.
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