Isolation and characterization of microsatellite loci in the finless porpoise (Neophocaena phocaenoides)

Eight dinucleotide microsatellite loci were isolated from the finless porpoise (Neophocaena phocaenoides). Analysis of 30 individuals showed the number of alleles ranged from four to 21 with observed heterozygosity ranging from 0.300 to 0.833, and expected heterozygosity ranging from 0.437 to 0.932. Cross‐species amplification was tested in four other cetacean species. These microsatellite markers would be valuable tools for population genetic studies of finless porpoises and other cetacean species.     

Development of tetranucleotide microsatellite loci for the finless porpoise (<Emphasis Type="Italic">Neophocaena phocaenoides</Emphasis>)

The finless porpoise, Neophocaena phocaenoides, is endemic to the coastal waters of the Indo-Pacific, ranging from the Persian Gulf to Japan. Nine tetranucleotide microsatellite loci were isolated from the finless porpoise (Neophocaena phocaenoides). Polymorphism of each locus was assessed in 39 unrelated individuals from the Yellow Sea and the South China Sea of the Chinese waters. The number of alleles per locus varied from 2 to 11. The ranges of observed and expected heterozygosity were 0.154–0.795 and 0.146–0.839, respectively. Cross-species amplification of these loci was tested in other cetacean species. These microsatellite markers described here will be suitable for population genetic studies of finless porpoises and other cetacean species.     

BILATERAL DIRECTIONAL ASYMMETRY OF THE APPENDICULAR SKELETON OF THE HARBOR PORPOISE (PHOCOENA PHOCOENA)

Directional asymmetry ( DA ) of the lengths, diameters, and masses of the scapula, humerus, radius, and ulna were analyzed on a sample of 213 harbor porpoises from Denmark and West Greenland. The levels of DA were consistent across yearlings and older animals, mature and immature animals, sexes, and populations. All investigated variables showed significant DA favoring the right side. For the pooled sample, DA of lengths ranged from 0.2% SE 0.1 (ulna) to 1.2% SE 0.1 (scapula). DA s of diameters were 1.5% SE 0.2 for the humerus, 0.6% SE 0.1 for the radius, and 1.3% SE 0.2 for the ulna. DA of mass ranged from 2.8% SE 0.5 (humerus) to 4.3% SE 0.7 (ulna). The humerus and ulna had significantly larger mean diameter/ length ratios on the right side than the left, making them more robust. The large DA of scapula length indicates larger muscle mass associated with the right flipper, while the generally more robust right humerii and ulnae may be designed for higher levels of mechanical stress. These DA s and the examples of lateralized behavior recorded in cetaceans, point to the existence of lateralized use of the flippers at the population level in harbor porpoises and possibly other cetacean species.     

Identification and characterization of a tandem repeat in exon III of the dopamine receptor D4 (DRD4) gene in cetaceans

A large number of mammalian species harbor a tandem repeat in exon III of the gene encoding dopamine receptor D4 (DRD4), a receptor associated with cognitive functions. In this study, a DRD4 gene exon III tandem repeat from the order Cetacea was identified and characterized. Included in our study were samples from 10 white-beaked dolphins (Lagenorhynchus albirostris), 10 harbor porpoises (Phocoena phocoena), eight sperm whales (Physeter macrocephalus), and five minke whales (Balaenoptera acutorostrata). Using enzymatic amplification followed by sequencing of amplified fragments, a tandem repeat composed of 18-bp basic units was detected in all of these species. The tandem repeats in white-beaked dolphin and harbor porpoise were both monomorphic and consisted of 11 and 12 basic units, respectively. In contrast, the sperm whale harbored a polymorphic tandem repeat with size variants composed of three, four, and five basic units. Also the tandem repeat in minke whale was polymorphic; size variants composed of 6 or 11 basic units were found in this species. The consensus sequences of the basic units were identical in the closely related white-beaked dolphin and harbor porpoise, and these sequences differed by a maximum of two changes when compared to the remaining species. There was a high degree of similarity between the cetacean basic unit consensus sequences and those from members of the horse family and domestic cow, which also harbor a tandem repeat composed of 18-bp basic units in exon III of their DRD4 gene. Consequently, the 18-bp tandem repeat appears to have originated prior to the differentiation of hoofed mammals into odd-toed and even-toed ungulates. The composition of the tandem repeat in cetaceans differed markedly from that in primates, which is composed of 48-bp repeat basic units.     

High‐density areas for harbor porpoises (Phocoena phocoena) identified by satellite tracking

The population status of harbor porpoises has been of concern for several years, and the establishment of Marine Protected Areas (MPAs) has been suggested as a method to protect the harbor porpoise (Phocoena phocoena, Linneaus 1758) and other small cetaceans. In order to designate MPAs, high‐density areas for the species must be identified. Spatial distribution of small cetaceans is usually assessed from ship or aerial surveys. As a potentially more accurate alternative, this study examined the movements and area preferences of 64 harbor porpoises, satellite tagged between 1997 and 2007, in order to determine the distribution in the North Sea, the western Baltic, and the waters in between. Results show that harbor porpoises are not evenly distributed, but congregate in nine high‐density areas within the study area. Several of these areas are subject to significant seasonal variation. The study found no differences in the home range size of males and females, but immature harbor porpoises have larger home ranges than mature porpoises. The use of satellite telemetry for identifying areas of high harbor porpoise density can be of key importance when designating MPAs.     

SKIN DENSITY AND ITS INFLUENCE ON BUOYANCY IN THE MANATEE (TRICHECHUS MANATUS LATIROSTRIS), HARBOR PORPOISE (PHOCOENA PHOCOENA), AND BOTTLENOSE DOLPHIN (TURSIOPS TRUNCATUS)

This study investigated how skin contributes to buoyancy control in the Florida manatee ( Trichechus manatus latirostris ), harbor porpoise ( Phocoena phocoena ), and bottlenose dolphin ( Tursiops truncatus ). Manatees are shallow divers and control their position in the water column hydrostatically. The two cetaceans are relatively deep divers that control their buoyancy hydrodynamically. Although the cetacean skin had been hypothesized to lower total body density ( e. g. , Dearolf et al. 2000, Nowacek et al. 2001), its buoyant force had not been calculated. The density of manatee skin, and its contribution to buoyancy, was unknown. Skin densities of 27 manatees, five harbor porpoises, and five bottlenose dolphins were measured volumetrically. Skin mass and density were used to calculate buoyant force. Harbor porpoise (952 kg/m³) and bottlenose dolphin (969 kg/m³) skins were less dense than seawater, and added 9 and 25 N of positive buoyant force, respectively, to total body buoyancy. By contrast, manatee skin (1,121 kg/m³) contributed 56 N of negative buoyant force, which equaled 70% of the negative buoyant force of their dense, pachyosteosclerotic ribs. Calculation of buoyant forces of the skeleton, skin and lungs demonstrates that the manatee is positively buoyant at the surface and negatively buoyant at depths of less than 10 m.     

HEMATOLOGY VALUES OF WILD HARBOR PORPOISES (PHOCOENA PHOCOENA) FROM THE BAY OF FUNDY,CANADA

Clinical hematology values were determined for 29 harbor porpoises (Phocoena phocoena) released from herring weirs in the Bay of Fundy, Canada. Erythrocyte values exhibited narrow ranges, but there was a high degree of individual variability in counts of white blood cells. Total white cell counts ranged from 2.6 to 15.5 ± 10⁹/liter, with an overall mean of 6.5 ± 2.7 ± 10±/liter. There were significant differences among reproductive classes in mean values of total red blood cells, hematocrit (HCT), mean cell volume (MCV), mean cell hemoglobin (MCHC) and total monocyte count. Wild harbor porpoises had fewer white blood cells, lower relative total number of neutrophils and lymphocytes, and higher percentages of monocytes and eosinophils than reported in the literature for captive porpoises. Compared to published values for other odontocetes, the hemograms of harbor porpoises were most similar to those of Pacific white-sided dolphins (Lagenorhynchus acutus). These hematology data represent a baseline from free-ranging harbor porpoises that can be used as a reference for long-term monitoring of the health of this population and as a tool for rehabilitation facilities.     

Increase in parasite burden and associated pathology in harbour porpoises (Phocoena phocoena) in West Greenland

Knowledge about parasitism in harbour porpoises and their health status around Greenland is scarce. This study provides knowledge about the poorly studied cetacean in its rapidly changing environment. Parasites and pathological findings in 20 harbour porpoises (Phocoena phocoena) hunted in waters around West Greenland are presented. Carcasses were dissected, and parasitological, histological and bacteriological investigations were carried out. Protozoa (Sarcocystis sp.), Nematoda (Halocercus invaginatus, Stenurus minor, Anisakis simplex sensu lato (s.l.), Crassicauda sp.), Trematoda (Campula oblonga) and Cestoda (Phyllobothrium delphini, Monorygma grimaldii) were found. Parasitic infection of the peribullar cavity and lung with pseudaliid nematodes was found in most animals. Sixty per cent of the porpoises were infected with stomach worms, and trematodes were present in liver and pancreas of 90 and 30 % of the porpoises, respectively. Crassicauda sp. was isolated from perimuscular fascia in 45 % of the animals. This is the first record of tetraphyllidean merocercoids in harbour porpoises. M. grimaldii and P. delphini were found in blubber layer of 15 % and abdominal cavity of 50 % of the porpoises. Bronchopneumonia, gastritis, cholangitis, pancreatitis and panniculitis were almost exclusively associated with parasitic infection and usually mild. Compared with a previous study of Greenlandic porpoises from 1995, a significant increase in severity of parasitic infections and the emergence of new parasite species were observed, most likely associated with changes in diet, influenced by increasing sea temperatures and receding ice cover.     

Lampyridae (Coleoptera): A plethora of mollicute associations

Beetles (Coleoptera) harbor many species ofAcholeplasma andSpiroplasma (division Tenericutes, class Mollicutes). Mollicutes were isolated from guts and/or hemocoels of firefly beetles (Lampyridae) from the United States (Maryland and West Virginia), Ecuador, and Tobago. Firefly beetles were frequent hosts for the group XIV spiroplasma, isolated from Ellychnia corrusca, and the group XIX spiroplasma, isolated fromPhoturis spp. The most unusual feature of the firefly-mollicute association is the carriage of four Mycoplasma species. Recent phylogenetic studies indicate that these species are members of a clade that includes a vertebrate pathogen,Mycoplasma mycoides. The high rate of occurrence ofMycoplasma species (which are, otherwise, infrequent in insects) in lampyrid beetles suggests that the association is significant. The unusual light-producing physiology of lampyrids (which is dependent on large pools of energy) and the production of large amounts of cardenolides from cholesterol (a critical growth factor for many mollicutes) may favor colonization by mollicutes. Offprint requests to: K. J. Hackett.     

Occurrence of a monophasic strain of Salmonella group B isolated from cetaceans in England and Wales between 1990 and 2002

Between September 1990 and December 2002, 511 cetacean carcasses stranded or caught in commercial fisheries in England and Wales were examined post mortem . Salmonella group B was isolated from 60 of 279 (21.51%) harbour porpoises ( Phocoena phocoena ), predominantly from lung tissue. Forty-three of the Salmonella group B isolates were subsequently serotyped and all found to have the antigenic structure O4,12:a:-. The annual proportion of harbour porpoises testing positive for Salmonella O4,12:a:- increased significantly from 6% in the early 1990s to 27% after 1999. The cause(s) of the increasing prevalence of Salmonella O4,12:a:- are not known, but may reflect natural variation in the epidemiological cycle of Salmonella O4,12:a:- in harbour porpoises. The probability of isolating Salmonella O4,12:a:- from harbour porpoises increased with age, suggesting that the mode of transmission is principally horizontal. There appeared to be a weak degree of seasonality in the probability of isolating Salmonella O4,12:a:- with a low proportion of cases in the months of April and May. Based on pathological findings from infected carcasses, Salmonella O4,12:a:- may be part of the normal commensal flora of the lungs of cetaceans with the potential to act as an opportunistic pathogen.     

Evidence of exposure to Brucella sp. in harbor porpoises (Phocoena phocoena) from the Bay of Fundy, Canada

Novel strains of Brucella recently have been discovered in marine mammals. To investigate Brucella exposure and infection in a general population of cetaceans, blood and tissue samples were collected and analyzed from wild harbor porpoises (Phocoena phocoena) incidentally caught in fishing gear in the Bay of Fundy, Canada. Two of 170 (1.2%) animals had detectable antibodies against Brucella, but no organisms were isolated from genital swabs or tissues from 22 and 8 porpoises, respectively. Genetic analysis of inflamed testes from 20 animals yielded no amplification of Brucella DNA. This is the first evidence of exposure to Brucella in porpoises from the western North Atlantic, and the prevalence is much lower than documented for conspecifics from the eastern North Atlantic.     

Timing of epiphyseal development in the flipper skeleton of the harbour porpoise (Phocoena phocoena) as an indicator of paedomorphosis

Epiphyseal development was investigated on X‐rays of flippers from 158 harbour porpoises from Danish waters. Development followed a proximodistal pattern similar to what is known in other cetacean species. Ossification of epiphyses was rare in the phalanges of the first and fifth digits and in the more distal phalanges of the second, third and fourth digits. Along with the morphology of the first metacarpal and the more distal phalanges this suggested paedomorphosis relative to delphinids. Male and female porpoises showed similar progression of epiphyseal development until approximately the sixth year. From then on, female porpoises showed more progressed development than males. This suggests a higher level of paedomorphosis in the male porpoise. The mechanism behind phocoenid paedomorphosis seems to be progenesis, probably as an adaptation towards a high reproductive rate relative to the delphinids.     

Life history of harbor porpoises (Phocoena phocoena) in Scottish (UK) waters

Life history parameters were determined for stranded and bycaught harbor porpoises (Phocoena phocoena) from Scottish (UK) waters (1992–2005). Fetal growth rate was 84.4 mm/mo and mean size at birth was 76.4 cm (range 65–88 cm). Males and females had a similar range of body lengths (65–170 cm and 66–173 cm, respectively), although asymptotic lengths were higher in females than males (approximately 158 cm and 147 cm, respectively). Nonpregnant females were significantly lighter, in relation to their length than males. Maximum estimated age was 20 yr for both sexes. Age at sexual maturity (ASM) was estimated as 4.35 yr in females and 5.00 yr in males. Conception occurred mainly in July and August although reproductively active males were recorded during April to July. Gestation lasted 10–11 mo, with calving mainly between May and July. Lactating females were recorded during June to November, while small calves with solid food in their stomachs were found mainly during February to May. Estimated pregnancy rate (0.34–0.40) is lower than recorded elsewhere, but is likely underestimated due to the prevalence of mature females of poor health status in the sample. Nevertheless, cetacean strandings can be an essential source of data on demographic parameters.     

Population structure of nuclear and mitochondrial DNA variation among South American Burmeister’s porpoises (<Emphasis Type="Italic">Phocoena spinipinnis</Emphasis>)

Little is known about the biology of Burmeister’s porpoises (Phocoena spinipinnis), a small cetacean species endemic to South American waters. Information on stock structure, however, is urgently needed, as the species suffers from considerable mortality due to local fishery activities throughout its distribution range. Using mitochondrial control region sequences and 11 species-specific microsatellite loci, we assessed the genetic differentiation among 118 stranded, incidentally or directly-caught Burmeister’s porpoises from different localities in Peruvian, Chilean, and Argentine waters. F-statistics and Bayesian clustering analyses indicate a major population differentiation along the South American Pacific coast, separating Peruvian from both Chilean and Argentine individuals. Interestingly, this population boundary is consistent with the population structure found in another sympatrically-occurring cetacean species: the dusky dolphin (Lagenorhynchus obscurus). Given that vulnerability to local depletion for South American coastal porpoises and dolphins is probably highest in the Peruvian population (due to high exploitation levels and recurrent El Niño events), the genetic data reported here considerably strengthen the need for conservation efforts focused on regulation of catches in local waters. Moreover, we discuss possible genetic differentiation among Burmeister’s porpoises (i) from the Atlantic and Pacific Ocean and (ii) from different Peruvian harbors. Finally, cross-species amplifications suggest that our newly-developed microsatellite markers will be useful in population genetic studies in the five other extant porpoise species.     

FORAGING STRATEGIES OF SYMPATRIC KILLER WHALE (ORCINUS ORCA) POPULATIONS IN PRINCE WILLIAM SOUND, ALASKA

Killer whales ( Orcinus orca ) feed on a wide variety of fish, cephalopods, and marine mammals throughout their cosmopolitan range; however, the dietary breadth that characterizes the species is not reflected in all populations. Here, we present the findings of a 14-yr study of the diet and feeding habits of killer whales in Prince William Sound, Alaska. Two non-associating forms of killer whale, termed resident and transient (Bigg et al. 1987), were identified. All prey seen taken by transients were marine mammals, including harbor seals ( Phoca vitulina ), Dall's porpoises ( Phocoenoides dalli ), Steller sea lions ( Eumetopias jubatus ), and harbor porpoises ( Phocoena phocoena ). Resident killer whales appeared to prey principally on salmon ( Oncorhynchus spp.), preferring coho salmon ( O. kisutch ) over other, more abundant salmon species. Pacific herring ( Clupea pallasi ) and Pacific halibut ( Hippocampus stenolepis ) were also taken. Resident killer whales frequently were seen to interact in non-predatory ways with Steller sea lions and Dall's porpoises, while transients were not. Differences in the social organization and behavior of the resident and transient killer whales in Prince William Sound are discussed in the light of the dietary differences documented here.     

Diet composition and food consumption rate of harbor porpoises (Phocoena phocoena) in the western Baltic Sea

Stomach content composition and prey‐specific consumption rates of juvenile and adult harbor porpoises (Phocoena phocoena) were estimated from a data set including 339 stomachs collected over a 32 yr period (1980–2011) in the western Baltic Sea. The stomach contents were mainly hard parts of fish prey and in particular otoliths. The bias originating from differential residence time of otoliths in the stomachs was addressed by use of a recently developed approach. Atlantic cod and herring were the main prey of adults, constituting on average 70% of the diet mass. Juvenile porpoises also frequently consumed gobies. Here, the mass contribution by gobies was on average 25%, which was as much as cod. Other species such as whiting, sprat, eelpout, and sandeels were of minor importance for both juveniles and adults. The diet composition differed between years, quarters, and porpoise acquisition method. Yearly consumption rates for porpoises in the western Baltic Sea were obtained in three scenarios on the daily energy requirements of a porpoise in combination with an estimate including the 95% CLs of the porpoise population size. Cod of age groups 1 and 2 and intermediate‐sized herring suffered the highest predation from porpoises.     

REACTIONS OF CAPTIVE HARBOR PORPOISES (PHOCOENA PHOCOENA) TO PINGER-LIKE SOUNDS

Pingers on gill nets can reduce bycatch of harbor porpoises. If harbor porpoises habituate to pingers, the effect may be reduced or lost. Two captive harbor porpoises were exposed to three sound types. All sounds were in the frequency band from 100 kHz to 140 kHz, 200 ms long, and presented once per 4 s. The source level was 153 dB re 1 μPa RMS at 1 m. Each session consisted of a 10‐min presound, a 5‐min sound, and a 10‐min postsound period. Behavior was recorded on video and on dataloggers placed on the dorsal fin of one animal. The loggers recorded heart rate, swimming speed, dive duration, and depth. The animals responded most strongly to the initial presentations of a sound. Surface time decreased, the heart rate dropped below the normal bradycardia, and echolocation activity decreased. The reactions of both animals diminished rapidly in the following sessions. Should the waning of responsiveness apply to wild animals, porpoises may adapt to the sounds but still avoid nets, or the bycatch may increase after some time. The success of long‐term use of pingers may then depend on the variety of sounds and rates of exposure.     

Parasites of the Dall's porpoise (Phocoenoides dalli True)

The prevalences of three helminths, Campula oblonga, Halocercus dalli and Crassicauda sp., recovered from Dall's porpoises which were net-entrapped incidentally in the vicinity of the Western Aleutian Islands in the northwest Pacific are reported. Specimens of Campula oblonga were found within the bile ducts of 46% of 127 livers examined. The prevalence of hepatic trematodiasis increased with the age of the host. Pulmonary nodules associated with Halocercus dalli were noted in 71% of the Dall's porpoises. Adult H. dalli were recovered from the main stem bronchi of heavily infected lungs. Younger animals exhibited a relatively higher prevalence. Specimens of Crassicauda sp. were found within the main lactiferous canal of 69% of 29 mammary glands examined. The prevalence was highest in mature porpoises. Possible detrimental effects and the modes of transmission of the three species of parasites are also considered.     

Direct isolation of mycoplasmas and acholeplasmas from sera and kidneys of calves.

Kidneys and sera of calves from various farms and primary kidney tissue cultures were tested for mycoplasma and acholeplasma contamination. Altogether 24 strains belonging to Mycoplasma arginini and Acholeplasma axanthum were isolated from tissue cultures, kidneys and sera. The same species were detected from lungs and peribronchial lymph nodes of calves, together with A. laidlawii, A. modicum and M. bovirhinis species. There was a close correlation between the mycoplasma content of tissue cultures, sera and kidneys and the clinical picture observed in the farm, as well as between the quality of tissue cultures and the mycoplasma content of organs, sera and tissue cultures.     

HARBOR PORPOISE (PHOCOENA PHOCOENA) ABUNDANCE IN ALASKA: BRISTOL BAY TO SOUTHEAST ALASKA, 1991-1993

Between 1991 and 1993, Alaska harbor porpoise ( Phocoena phocoena ) abundance was investigated during aerial surveys throughout much of the coastal and offshore waters from Bristol Bay in the eastern Bering Sea to Dixon Entrance in Southeast Alaska. Line-transect methodology was used, and only those observations made during optimal conditions were analyzed. Survey data indicated densities of 4.48 groups/100 km², or approximately 3,531 harbor porpoises (95% C. I. 2,206-5,651) in Bristol Bay and 0.54 groups/100 km², or 136 harbor porpoises (95% C. I. 11-1,645) for Cook Inlet. Efforts off Kodiak Island resulted in densities of 1.85 groups/100 km², or an abundance estimate of 740 (95% C. I. 259-2,115). Surveys off the south side of the Alaska Peninsula found densities of 2.03 groups/100 km² and an abundance estimate of 551 (95% C. I. 423-719). Surveys of offshore waters from Prince William Sound to Dixon Entrance yielded densities of 4.02 groups/100 km² and an abundance estimate of 3,982 (95% C. I. 2,567-6,177). Combining all years and areas yielded an uncorrected density estimate of 3.82 porpoises per 100 km², resulting in an abundance estimate of 8,940 porpoises (CV = 13.8%) with a 95% confidence interval of 6,746-11,848. Using correction factors from other studies to adjust for animals missed by observers, the total number of Alaska harbor porpoises is probably three times this number.