A tradeoff evolution between acoustic fat bodies and skull muscles in toothed whales.

Toothed whales have developed specialized echolocation abilities that are crucial for underwater activities. Acoustic fat bodies, including the melon, extramandibular fat body, and intramandibular fat body, are vital for echolocation. This study explores the transcriptome of acoustic fat bodies in toothed whales, revealing some insight into their evolutionary origins and ecological significance. Comparative transcriptome analysis of acoustic fat bodies and related tissues in a harbor porpoise and a Pacific white-sided dolphin reveals that acoustic fat bodies possess characteristics of both muscle and adipose tissue, occupying an intermediate position. The melon and extramandibular fat body exhibit specific muscle-related functions, implying an evolutionary connection between acoustic fat bodies and muscle tissue. Furthermore, we suggested that the melon and extramandibular fat body originate from intramuscular adipose tissue, a component of white adipose tissue. The extramandibular fat body has been identified as an evolutionary homolog of the masseter muscle, supported by the specific expression of MYH16, a pivotal protein in masticatory muscles. The intramandibular fat body, located within the mandibular foramen, shows possibilities of the presence of several immune-related functions, likely due to its proximity to bone marrow. Furthermore, this study sheds light on leucine modification in the catabolic pathway, which leads to the accumulation of isovaleric acid in acoustic fat bodies. Swallowing without chewing, a major toothed whale feeding ecology adaptation, makes the masticatory muscle redundant and leads to the formation of the extramandibular fat body. We propose that the intramuscular fat enlargement in facial muscles, which influences acoustic fat body development, is potentially related to the substantial reorganization of head morphology in toothed whales during aquatic adaptation.

Notes on stomach contents of pygmy and dwarf sperm whales (Kogia spp.) from around Japan.

The diets of pygmy (Kogia breviceps) and dwarf (K. sima) sperm whales in Japanese waters are poorly known. We report new information on the diets of these two species from these waters based on identifiable hard-part remains recovered from the stomach contents of 29 whales (11 pygmy and 18 dwarf sperm whales) that stranded between 1991 and 2021; those of a further two dwarf sperm whales were empty. The cephalopod (and secondarily fish and crustacean) component of the diets of these 29 whales, based on analysis of identifiable stomach-content remains, is described. The main prey includes cephalopods, represented by 1556 identifiable lower beaks (and 1483 upper beaks), crustaceans (represented by heavily digested, unidentifiable remains), and fishes (as represented by 92 otoliths). Identified prey comprises 30 species from 16 cephalopod families and 5 families from 5 fish orders. Oceanic cephalopods are the main prey of both whale species, particularly Enoploteuthis (Paraenoploteuthis) chunii and Chiroteuthis (Chirothauma) picteti. Prey diversity index values (Shannon-Weaver's diversity index H') are 2.41 for the pygmy sperm whale and 2.66 for the dwarf sperm whale. Although the main cephalopod component in the diets of these two whale species is similar, Pianka's index (0.40), a measure of niche overlap, is not that high, and may be influenced by differences in prey dominance in different feeding areas.

Assessing the costs of GHG emissions of multi-product agricultural systems in Vietnam.

Besides a vital sector of the economy, agriculture is a primary source of greenhouse gas (GHG) emissions. The present paper investigates the impact of carbon tax policy on Vietnamese agriculture by focusing on multi-product systems such as rice, livestock, and aquaculture, traditionally called the Vuon (Garden)-Ao (Pond)-Chuong (livestock pen) system (VAC). In it, farmers use garden, pond, and pen by-products as fertilizer and feed. We use shadow prices and Morishima substitution elasticities as greenhouse gas emissions indicators, estimated with directional output distance function. Farmers in the Mekong Delta region are found to be technologically less efficient than in other regions of Vietnam, though the shadow prices of GHG emissions are lower there too. This indicates that farmers in the Mekong Delta, generally concentrating either on livestock or aquaculture, have greater potential for reducing GHG emissions by way of improvements in technical efficiency than do those in other regions. However, Morishima elasticity estimates show that policy impacts diminish more quickly in the Mekong than elswhere. We suggest the Vietnamese government encourage Mekong Delta farmers to employ technologically more efficient methods or shift to more balanced farming to reduce the shadow price of GHG emissions, encouraging more efficient emissions reduction.

Diphyllobothrium stemmacephalum (Cestoda: Diphyllobothriidae) found from a harbor porpoise in northern Japan, with comments on a geographical gap with human infection cases in southern Japan.

Even though the cetacean tapeworm Diphyllobothrium stemmacephalum occurs in both cold and warm waters, human infections and final host occurrences have been confined to temperate areas in and near Japan. We recently obtained a strobila of this cestode that was excreted from a harbor porpoise accidentally caught offshore of Hokkaido of northern Japan. Genetic analysis of 28S rDNA and cox1 genes confirmed that the cestode was D. stemmacephalum. Our finding sets the northernmost record of D. stemmacephalum in the western Pacific, suggesting that the risk of human infections by this parasite in northern Japan deserves further attention.

Gastric ulceration caused by genetically identified Anisakis simplex sensu stricto in a harbor porpoise from the Western Pacific stock.

The genus Anisakis is a well-known group of nematodes that parasitize cetaceans as the final host and cause mucosal damage to their stomach. However, little has been done to precisely identify the nematodes recovered from the final hosts, especially in the Western Pacific, because of taxonomic confusion about the discrimination of sibling species and the difficulties of obtaining specimens from cetaceans. We describe the results of genetic identification and histopathological observations of specimens recovered from an ulcerated lesion and stomach contents in the forestomach of a female harbor porpoise accidentally caught by a set net fishery in Usujiri, southern Hokkaido, Japan. All the specimens arbitrarily collected from the lesion and stomach contents were identified as Anisakis simplex sensu stricto according to their ITS rDNA sequences. The size of the ulcer was approximately 6.3 mm in diameter and it was infected with 119 individual nematodes, mostly consisting of L3 and L4 stage larvae (95.0%). Histological sections were characterized by a locally extensive ulcer with the parasites penetrating into the muscularis externa that caused a thickening of the surrounding mucosa.

Foraging activity of harbour porpoises around a bottom-gillnet in a coastal fishing ground, under the risk of bycatch.

Bycatch of harbour porpoises (Phocoena phocoena) by gillnets is a recognised threat to populations. To develop effective mitigation measures, understanding the mechanics of bycatch is essential. Previous studies in experimental conditions suggested foraging activity is an important factor influencing porpoises' reaction to gillnets. We acoustically observed the behaviour of wild harbour porpoises around a bottom-gillnet set-up in a commercial fishing ground, especially foraging activity. Passive acoustic event recorders (A-tags) were fixed to the ends of the gillnet, and recorded for 1 392 hours. Although harbour porpoises frequently and repeatedly appeared around the net each day, incidental bycatch occurred only three times during the observations. The stomach contents of two individuals contained mainly Ammodytes sp., which were observable around the bottom-gillnet but not targeted by the fishery. A total of 276 foraging incidents were acoustically detected, and 78.2% of the foraging activity was in the bottom layer (deeper than 25 m). Porpoises appeared around the net with more frequency on the day of a bycatch incident than on the days without bycatch. These results suggest that the harbour porpoises appeared around the bottom-gillnet to forage on fish distributed in the fishing ground, but not captured by this bottom-gillnet. Thus, porpoises face the risk of becoming entangled when foraging near a gillnet, with the probability of bycatch simply increasing with the length of time spent near the net. Bycatch mitigation measures are discussed.

Serologic survey of Brucella infection in cetaceans inhabiting along the coast of Japan.

A serologic investigation of Brucella infection was performed in 7 species of cetaceans inhabiting along the coast of Japan. A total of 32 serum samples were examined by enzyme-linked immunosorbent assay (ELISA) using Brucella abortus and B. canis antigens. One serum sample from five melon-headed whales (Peponocephala electra) was positive for B. abortus. No serum sample showed positive for B. canis. The ELISA-positive melon-headed whale serum demonstrated a strong band appearance only against B. abortus antigens in Western blot analysis. Many detected bands were discrete, while some of them had a smeared appearance. The present results indicate that Brucella infection occurred in melon-headed whale population and the bacterial antigenicity is more similar to that of B. abortus than B. canis.

Description of a new species of beaked whale (Berardius) found in the North Pacific.

Two types of Berardius are recognised by local whalers in Hokkaido, Japan. The first is the ordinary Baird's beaked whale, B. bairdii, whereas the other is much smaller and entirely black. Previous molecular phylogenetic analyses revealed that the black type is one recognisable taxonomic unit within the Berardius clade but is distinct from the two known Berardius species. To determine the characteristics of the black type, we summarised external morphology and skull osteometric data obtained from four individuals, which included three individuals from Hokkaido and one additional individual from the United States National Museum of Natural History collection. The whales differed from all of their congeners by having the following unique characters: a substantially smaller body size of physically mature individuals, proportionately shorter beak, and darker body colour. Thus, we conclude that the whales are a third Berardius species.

Surveillance of amyloidosis in stranded and bycaught cetaceans off Hokkaido, Japan.

Systemic amyloidosis is rarely reported among cetaceans, and a surveillance dedicated for its occurrence across a certain geographic location has not been reported. Between 2013 and 2018, comprehensive gross and histopathologic examinations were conducted on 54 animals comprising 11 species of stranded and bycaught cetaceans in Hokkaido, Japan. Systemic amyloidosis was diagnosed in 2 out of 3 Stejneger's beaked whales (Mesoplodon stejnegeri), through Congo red staining and immunohistochemistry for amyloid A. The kidney and gastrointestinal tract had the largest amounts of amyloid deposits, representing a previously undescribed organ distribution in the species. The current study demonstrates the possibility of Stejneger's beaked whales being prone to the development of systemic amyloidosis, and highlights the need for further investigations.

Different profiles of naturally produced and anthropogenic organohalogens in the livers of cetaceans from the Sea of Japan and the North Pacific Ocean.

Levels and profiles of naturally produced halogenated bipyrroles (Br4Cl2-DBP and Cl7-MBP), methoxylated tetrabromodiphenyl ethers (6-MeO-BDE47), anthropogenic perfluoroalkyl substances (PFASs) and legacy persistent organic pollutants (POPs) were investigated in the livers of 14 cetaceans from the Sea of Japan and the North Pacific Ocean. The concentrations of Br4Cl2-DBP (4 to 4900 ng/g-wet), Cl7-MBP (16 to 3960 ng/g-wet) and 6-MeO-BDE47 (7 to 190 ng/g-wet) were higher in the order of killer whales > toothed whales > baleen whales. Profiles of PFASs were dominated by perfluoroundecanoic and perfluorotridecanoic acids (10 to 540 ng/g-wet), sum of which accounted for 70% of total measured PFASs. Regional difference was observed for Cl7-MBP and PFASs, which were higher in the Sea of Japan, whereas Br4Cl2-DBP was in the North Pacific Ocean. Specific accumulation pattern of these natural contaminants in cetaceans around northern Japan could help compare the exposure profile of PFASs and POPs among other geographic regions.

Click emission in Dall's porpoise Phocoenoides dalli, focusing on physical properties of tissues.

Dall's porpoise (Phocoenoides dalli) is one of most common North Pacific porpoise species, for which information on sound-emitting processes is limited. To evaluate the mechanism of click emission in the head of this porpoise, the distribution of acoustic impedance in head tissues was calculated using density and Young's modulus'which is a measure of linear resistance to linear compression. Two Dall's porpoise heads were examined: one for macroscopic dissection, and one for investigating the distribution of acoustic impedance calculated from CT-measured density, and Young's modulus measured by creep meter. Acoustic impedance increased from the dorsal bursae to the melon's emitting surface, with impedance matching observed at the boundary between the emitting surface and seawater, and was more similar in distribution to Young's modulus than it was to density. The distribution of acoustic impedance differed from that of harbor porpoise (Phocoena phocoena), despite similarities in the sound-producing organs in the heads of Dall's and harbor porpoises. A comparison of the physical properties of Dall's and harbor porpoise head tissues suggests that hypertrophic vestibular sacs and an oval emitting surface are common characteristics in Phocoenidae.

Prominent hepatic ductular reaction induced by Oschmarinella macrorchis in a Hubbs' beaked whale Mesoplodon carlhubbsi, with biological notes.

Beaked whales are among the least known group of cetaceans, and information regarding their pathology and parasitology is especially scarce. We describe a case of significant parasitism by a trematode found in the liver of an adult male Hubbs' beaked whale Mesoplodon carlhubbsi that stranded in Hokkaido, Japan. Post-mortem examinations revealed a localised area of discolouration restricted to the hilar region of the left hepatic lobe, where spindle-shaped trematodes occupied the dilated and hypertrophic bile ducts. Histologically, the intrahepatic bile ducts were characterised by adenomatous hyperplasia with goblet cell metaplasia of the biliary epithelium. Findings in the adjacent hepatic parenchyma included pseudocarcinomatous ductular reactions obliterating hepatocytes, a histomorphology not previously reported in marine mammals. Morphological identification of the trematode corresponded to Oschmarinella macrorchis, which has only been reported once in a Stejneger's beaked whale, M. stejnegeri. PCR amplification and sequencing analyses of the parasite's mtDNA ND3, 18S and 28S rRNA regions generated novel gene sequences. Environmental contaminant levels were measured to explore its potential relationship with the parasitism but there was no conclusive association. A high level of polychlorinated biphenyl (30000 ng g-1 lipid weight) was detected in the blubber of this individual, when compared to those of 3 other male Hubbs' beaked whales stranded in Japan. Stomach contents were also analysed, indicating the presence of various squid species and unidentified fish. Our results contribute to the knowledge of a little-known beaked whale and provide evidence for the first time of the pathobiological response caused by O. macrorchis.

Tissue-specific bioaccumulation of long-chain perfluorinated carboxylic acids and halogenated methylbipyrroles in Dall's porpoises (Phocoenoides dalli) and harbor porpoises (Phocoena phocoena) stranded in northern Japan.

This study investigated accumulation of perfluoroalkyl substances (PFASs), persistent organochlorines (OCs), and naturally produced halogenated compounds (NHCs), including brominated methylbipyrroles and methoxylated bromodiphenyl ethers, in liver, blood, and blubber from Dall's porpoises (Phocoenoides dalli) and harbor porpoises (Phocoena phocoena) stranded in Hokkaido, northern Japan. Profiles of the PFASs were dominated by perfluoroundecanoic acid and perfluorotridecanoic acid, both of which accounted for 70% of the total measured PFAS concentrations in both porpoise species. The mean concentrations of the ∑PFCA were 573ng/g wet weight (ng/g-wet) in liver, 62ng/g-wet in whole blood, and 28ng/g-wet in blubber from the Dall's porpoises, and were significantly higher (p<0.05) than those in the harbor porpoises. The hepatic concentrations of perfluorooctane sulfonate (PFOS) were <14ng/g-wet, and accounted for only 3% of the total measured PFASs. The profiles of PFASs in the porpoises resembled those in fish species in this area, implying a common source of exposure to PFASs in East Asia. On the other hand, in the blubber of Dall's porpoises, NHCs were dominated by 2,3,3',4,4',5,5'-heptachloro-1'-methyl-1,2'-bipyrrole (867ng/g-wet), 5,5'-dichloro-1,1'-dimethyl-3,3',4,4'-tetrabromo-2,2'-bipyrrole (481ng/g-wet), and 6-methoxy-2,2',4,4'-tetrabromodiphenyl ether (30ng/g-wet), which were present at higher concentrations than in harbor porpoises. Factor analysis with varimax rotation revealed that factor 1 had higher eigenvectors (element in eigenvalues) for long-chain PFCAs and PFOS, which was found in the highest concentrations in the liver, whereas factor 2 was mainly associated with lipid soluble NHCs and OCs in both species. No correlations were observed between long-chain PFCAs and NHCs in the porpoises, probably because of the different sources and accumulation kinetics. Future research should assess the temporal trends and long-term effects of PFASs and NHCs in the tissues of mammals from the Asia-Pacific region.

Description of the karyotypes of Stejneger's beaked whale (Mesoplodon stejnegeri) and Hubbs' beaked whale (M. carlhubbsi)

Abstract The genus Mesoplodon (Cetacea: Odontoceti: Ziphiidae) is one of the few cetacean genera with the karyotype 2n = 42. The 2n = 42 karyotype of M. europaeus and M. carlhubbsi is largely consistent with the general cetacean karyotype 2n = 44, although other 2n = 42 karyotypes do not exhibit clear homologies with the general cetacean karyotype. Therefore, the chromosomes of Mesoplodon species may be the key to understanding cetacean karyological evolution. In the present study, the male karyotypes of M. stejnegeri and M. carlhubbsi were examined. In both species, the diploid number of the male karyotype was 42. Both species had the following characteristics: 1) a huge subtelocentric X chromosome with a large C-block; 2) a small metacentric Y chromosome; 3) nucleolus organizer regions (NORs) in the terminal regions of a large autosome and one or two small metacentric autosomes; 4) small metacentric autosomes; 5) large submetacentric and subtelocentric autosomes; 6) less accumulated C-heterochromatin in the centromeric region; and 7) heteromorphism in C-heterochromatin accumulation between homologues. Characteristics 1 and 3 are peculiar to only the karyotypes of Mesoplodon species, whereas characteristics 4, 5, 6, and 7 are also found in the species with the general cetacean karyotype 2n = 44.

Description of the karyotypes of Stejneger's beaked whale (Mesoplodon stejnegeri) and Hubbs' beaked whale (M. carlhubbsi).

The genus Mesoplodon (Cetacea: Odontoceti: Ziphiidae) is one of the few cetacean genera with the karyotype 2n = 42. The 2n = 42 karyotype of M. europaeus and M. carlhubbsi is largely consistent with the general cetacean karyotype 2n = 44, although other 2n = 42 karyotypes do not exhibit clear homologies with the general cetacean karyotype. Therefore, the chromosomes of Mesoplodon species may be the key to understanding cetacean karyological evolution. In the present study, the male karyotypes of M. stejnegeri and M. carlhubbsi were examined. In both species, the diploid number of the male karyotype was 42. Both species had the following characteristics: 1) a huge subtelocentric X chromosome with a large C-block; 2) a small metacentric Y chromosome; 3) nucleolus organizer regions (NORs) in the terminal regions of a large autosome and one or two small metacentric autosomes; 4) small metacentric autosomes; 5) large submetacentric and subtelocentric autosomes; 6) less accumulated C-heterochromatin in the centromeric region; and 7) heteromorphism in C-heterochromatin accumulation between homologues. Characteristics 1 and 3 are peculiar to only the karyotypes of Mesoplodon species, whereas characteristics 4, 5, 6, and 7 are also found in the species with the general cetacean karyotype 2n = 44.

Polybrominated diphenyl ethers (PBDEs) and their hydroxylated and methoxylated analogues in the blood of harbor, Dall's and finless porpoises from the Japanese coastal waters.

This study investigated the accumulation of polybrominated diphenyl ethers (PBDEs) and their hydroxylated and methoxylated analogues (OH-PBDEs and MeO-PBDEs) in the blood of harbor porpoises, Dall's porpoises, and finless porpoises stranded or bycaught in Japanese coastal waters and in the North Pacific Ocean. Moreover, we suggested the origins of these contaminants and the factors affecting their pattern of accumulation. Levels of PBDEs in Dall's porpoises were one order of magnitude greater than those in the other species. OH-PBDE and MeO-PBDE levels were comparable to those of PBDEs. However, no correlation was found between the levels of OH-PBDEs and PBDEs, whereas a strong correlation was found between that of OH-PBDEs and MeO-PBDEs (p < 0.001). 6OH-BDE47, reported compound biosynthesized by marine low-trophic level organisms, was the dominant congener. These results suggest that PBDEs found in these porpoise species derive from flame retardants, but OH-PBDEs and MeO-PBDEs are mainly of natural origins.

Concentration and congener pattern of polychlorinated biphenyls in blubber and liver of Hubbs' beaked whale (Mesoplodon carlhubbsi), using a sulfoxide and an Ag-ION solid phase extraction cartridge as a simplified cleanup technique for biological samples.

We performed the first known study of polychlorinated biphenyls (PCBs) concentrations and patterns in the blubber and liver of a Hubbs' beaked whale. Samples were pretreated with Supelclean™ sulfoxide and Discovery® Ag-ION solid phase extraction cartridges to remove whale oil. PCB concentrations in the blubber and liver were 13,000 and 7300ng/g lipid, respectively. Highly poisonous congeners such as dioxin-like (DL) PCBs tended to accumulate in the liver. The toxic equivalents (TEQ) of DL-PCBs in the liver (740pg-TEQ/g lipid) were higher than those in the blubber (74pg-TEQ/g lipid). The blubber and liver samples showed that hexachlorinated biphenyls were dominant among homologues, and PCB-153 was dominant among congeners. Several congeners accumulated disproportionately in the blubber and the liver (PCB-28, 52, 74, 99, and 118), while others did not persist (PCB-31, 70, and 110). This indicates that PCBs are metabolized differently according to their specific composition.

Tissue physical property of the harbor porpoise Phocoena phocoena for investigation of the sound emission process.

The process by which sound is propagated in the head of a toothed whale is still a subject of discussion. Investigating the distribution of acoustic impedance calculated by density and Young's modulus is effective for quantitative comprehension because acoustic impedance determines the reflection coefficient of a sound wave. However, the sound propagation process of the toothed whale has been mainly examined by either anatomical techniques or the measurement of density or sound velocity. In the current study, the acoustic impedance of head tissue of harbor porpoise was measured. Results of this study should be a helpful information for further discussion about the relationship between the structure of sound-producing organ and clicks property.

Brown adipose tissue in cetacean blubber.

Brown adipose tissue (BAT) plays an important role in thermoregulation in species living in cold environments, given heat can be generated from its chemical energy reserves. Here we investigate the existence of BAT in blubber in four species of delphinoid cetacean, the Pacific white-sided and bottlenose dolphins, Lagenorhynchus obliquidens and Tursiops truncates, and Dall's and harbour porpoises, Phocoenoides dalli and Phocoena phocoena. Histology revealed adipocytes with small unilocular fat droplets and a large eosinophilic cytoplasm intermingled with connective tissue in the innermost layers of blubber. Chemistry revealed a brown adipocyte-specific mitochondrial protein, uncoupling protein 1 (UCP1), within these same adipocytes, but not those distributed elsewhere throughout the blubber. Western blot analysis of extracts from the inner blubber layer confirmed that the immunohistochemical positive reaction was specific to UCP1 and that this adipose tissue was BAT. To better understand the distribution of BAT throughout the entire cetacean body, cadavers were subjected to computed tomography (CT) scanning. Resulting imagery, coupled with histological corroboration of fine tissue structure, revealed adipocytes intermingled with connective tissue in the lowest layer of blubber were distributed within a thin, highly dense layer that extended the length of the body, with the exception of the rostrum, fin and fluke regions. As such, we describe BAT effectively enveloping the cetacean body. Our results suggest that delphinoid blubber could serve a role additional to those frequently attributed to it: simple insulation blanket, energy storage, hydrodynamic streamlining or contributor to positive buoyancy. We believe delphinoid BAT might also function like an electric blanket, enabling animals to frequent waters cooler than blubber as an insulator alone might otherwise allow an animal to withstand, or allow animals to maintain body temperature in cool waters during sustained periods of physical inactivity.

Stable isotope ratios of carbon, nitrogen and oxygen in killer whales (Orcinus orca) stranded on the coast of Hokkaido, Japan.

We analyzed δ(13)C, δ(15)N and δ(18)O in the muscle and liver from killer whales stranded on the coast of Japan. The δ(15)N values in the muscle samples from calves were apparently higher than those in their lactating mothers, suggesting that nursing may result in the higher δ(15)N values in the muscle samples of calves. The δ(15)N value in the muscle samples of male and female whales, except for the calves, were positively correlated with the δ(13)C values and body length, suggesting that the increases in δ(15)N were due to the growth of the whales and increase in their trophic level. In contrast, the δ(18)O values in the muscle samples of female whales except for the calves were negatively correlated with the δ(13)C and δ(15)N values. The δ(18)O may be lower in whales occupying higher trophic positions (δ(15)N), although it might also be affected by geographic and climatic conditions.