Isolation and genetic characterization of Toxoplasma gondii from striped dolphin (Stenella coeruleoalba) from Costa Rica

Toxoplasma gondii infection in marine mammals is of interest because of mortality and mode of transmission. It has been suggested that marine mammals become infected with T. gondii oocysts washed from land to the sea. We report the isolation and genetic characterization of viable T. gondii from a striped dolphin (Stenella coeruleoalba), the first time from this host. An adult female dolphin was found stranded on the Pacific Coast of Costa Rica, and the animal died the next day. The dolphin had a high (1:6400) antibody titer to T. gondii in the modified agglutination test. Severe nonsuppurative meningoencephalomyelitis was found in its brain and spinal cord, but T. gondii was not found in histological sections of the dolphin. Portions of its brain and the heart were bioassayed in mice for the isolation of T. gondii. Viable T. gondii was isolated from the brain, but not from the heart, of the dolphin. A cat fed mice infected with the dolphin isolate (designated TgSdCol) shed oocysts. Genomic DNA from tachyzoites of this isolate was used for genotyping at 10 genetic loci, including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico, and this TgSdCo1 isolate was found to be Type II.     

A new species of the genus Zalophotrema (Digenea: Campulidae), Zalophotrema atlanticum n. sp., from the liver of the striped dolphin Stenella coeruleoalba (Meyen, 1833) (Cetacea: Delphinidae) in Atlantic waters

Zalophotrema atlanticum n. sp. from the liver of a striped dolphin, Stenella coeruleoalba (Meyen, 1833), stranded in the Canary Islands is described. This new species differs from the other Zalophotrema species in body dimensions and relative size and position of the suckers and cirrus-sac. This is the first record of a species of the genus Zalophotrema in the striped dolphin, and the Atlantic Ocean constitutes a new locality record for this genus.     

Herpes simplex-like infection in a bottlenose dolphin stranded in the Canary Islands

A bottlenose dolphin, stranded in the Canary Islands in 2001 exhibited non-suppurative encephalitis. No molecular detection of cetacean morbillivirus (CeMV) was found, but a herpesviral-specific band of 250 bp was detected in the lung and brain. The sequenced herpesviral PCR product was compared with GenBank sequences, obtaining 98% homology (p-distance of 0.02) with Human herpesvirus 1 (herpes simplex virus 1 or HSV-1). This is the first report of a herpes simplex-like infection in a stranded dolphin.     

Hearing loss in stranded odontocete dolphins and whales

The causes of dolphin and whale stranding can often be difficult to determine. Because toothed whales rely on echolocation for orientation and feeding, hearing deficits could lead to stranding. We report on the results of auditory evoked potential measurements from eight species of odontocete cetaceans that were found stranded or severely entangled in fishing gear during the period 2004 through 2009. Approximately 57% of the bottlenose dolphins and 36% of the rough-toothed dolphins had significant hearing deficits with a reduction in sensitivity equivalent to severe (70-90 dB) or profound (>90 dB) hearing loss in humans. The only stranded short-finned pilot whale examined had profound hearing loss. No impairments were detected in seven Risso's dolphins from three different stranding events, two pygmy killer whales, one Atlantic spotted dolphin, one spinner dolphin, or a juvenile Gervais' beaked whale. Hearing impairment could play a significant role in some cetacean stranding events, and the hearing of all cetaceans in rehabilitation should be tested.     

Inter-species differences in polychlorinated biphenyls patterns from five sympatric species of odontocetes: Can PCBs be used as tracers of feeding ecology?

Concentrations of thirty two polychlorinated biphenyls (PCBs) were determined in the blubber of five sympatric species of odontocetes stranded or by-caught along the Northwest coast of the Iberian Peninsula: common dolphin (Delphinus delphis), long-finned pilot whale (Globicephala melas), harbour porpoise (Phocoena phocoena), striped dolphin (Stenella coeruleoalba) and bottlenose dolphin (Tursiops truncatus). Multivariate analyses were applied to evaluate the ability of PCB patterns to discriminate these sympatric species and to determine which eco-biological factors influence these patterns, thus evaluating the relevance of PCB concentrations as biogeochemical tracers of feeding ecology. The five species could be separated according to their PCB patterns. Different exposure to these contaminants, a consequence of their different dietary preferences or habitats, together with potentially dissimilar metabolic capacities, likely explain these results; sex, age, habitat and the type of prey eaten were the most important eco-biological parameters of those tested. Although, no single congener has been specifically identified as a tracer of feeding ecology, 4 congeners from the 22 analysed seemed to be the most useful and around 12 congeners appear to be enough to achieve good discrimination of the cetaceans studied. Therefore, this study suggests that PCB patterns can be used as tracers for studying the feeding ecology, sources of contamination or even population structure of cetacean species from the Northwest Iberian Peninsula.     

MORBILLIVIRUS INFECTION IN MANATEES

The first evidence of exposure of free-ranging Florida manatees ( Trichechus manatees latirostris ) to a morbillivirus is reported. Blood samples were collected from 148 Florida manatees between 1977 and 1994. The sample included manatees that were under rehabilitation (n = 97), free-ranging (n = 40) and fresh necropsy specimens (n = 11). Serum from six animals (4%) neutralized porpoise and dolphin morbilliviruses to a higher titer than any other member of the Morbillivirus genus. Antibodies were not detected in sera from 12 free-ranging Antillean manatees ( T. manatus manatus ) from Guyana sampled in 1992 or from 12 hand-reared Amazonian manatees ( T. inunguis ) sampled in 1984. Immunoprecipitation studies using radio-labelled canine distemper virus protein and serum from Florida manatees showed precipitation of the nucleo-capsid (N) protein. The combination of low antibody titers and absence of clinical disease suggest that the Florida manatee is a dead-end host. Sporadic infection may occur following contact with another species in which infection is enzootic. Morbillivirus could, either by fatal infection or more insidious effects on the immune system or reproduction, pose a problem to this already threatened species.     

Seroprevalence of Toxoplasma gondii antibodies in wild dolphins from the Spanish Mediterranean coast

Although Toxoplasma gondii infection has been found occasionally in cetaceans, little is known of the prevalence of antibodies to T. gondii in wild dolphins. Antibodies to T. gondii were determined in serum samples from 58 dolphins stranded in the Spanish Mediterranean coast. Modified agglutination test was used to determine T. gondii antibodies, and a titer of 1:25 was considered indicative of T. gondii infection. Antibodies to T. gondii were found in 4 of 36 striped dolphins (Stenella coeruleoalba), in 2 of 4 common dolphins (Delphinus delphis), in 4 of 7 bottlenose dolphins (Tursiops truncatus), and in 1 harbour porpoise (Phocoena phocoena). Antibodies were not found in 9 Risso's dolphins (Grampus griseus) and in 1 long-finned pilot whale (Globicephala melas) surveyed. The results indicate that T. gondii infection is frequent in at least 3 dolphin species from the Mediterranean Sea.     

Demographic Clusters Identified within the Northern Gulf of Mexico Common Bottlenose Dolphin (Tursiops truncates) Unusual Mortality Event: January 2010 - June 2013

A multi-year unusual mortality event (UME) involving primarily common bottlenose dolphins (Tursiops truncates) was declared in the northern Gulf of Mexico (GoM) with an initial start date of February 2010 and remains ongoing as of August 2014. To examine potential changing characteristics of the UME over time, we compared the number and demographics of dolphin strandings from January 2010 through June 2013 across the entire GoM as well as against baseline (1990-2009) GoM stranding patterns. Years 2010 and 2011 had the highest annual number of stranded dolphins since Louisiana’s record began, and 2011 was one of the years with the highest strandings for both Mississippi and Alabama. Statewide, annual numbers of stranded dolphins were not elevated for GoM coasts of Florida or Texas during the UME period. Demographic, spatial, and temporal clusters identified within this UME included increased strandings in northern coastal Louisiana and Mississippi (March-May 2010); Barataria Bay, Louisiana (August 2010-December 2011); Mississippi and Alabama (2011, including a high prevalence and number of stranded perinates); and multiple GoM states during early 2013. While the causes of the GoM UME have not been determined, the location and magnitude of dolphin strandings during and the year following the 2010 Deepwater Horizon oil spill, including the Barataria Bay cluster from August 2010 to December 2011, overlap in time and space with locations that received heavy and prolonged oiling. There are, however, multiple known causes of previous GoM dolphin UMEs, including brevetoxicosis and dolphin morbillivirus. Additionally, increased dolphin strandings occurred in northern Louisiana and Mississippi before the Deepwater Horizon oil spill. Identification of spatial, temporal, and demographic clusters within the UME suggest that this mortality event may involve different contributing factors varying by location, time, and bottlenose dolphin populations that will be better discerned by incorporating diagnostic information, including histopathology.     

Fatal Asphyxiation in Bottlenose Dolphins (Tursiops truncatus) from the Indian River Lagoon

Multiple single case reports of asphyxiation in dolphins caused by fish lodged in the esophagus exist. However, the significance of this cause of mortality in a single population has not been documented. We performed a retrospective evaluation of pathology records from stranded bottlenose dolphins (Tursiops truncatus) from the Indian River Lagoon to evaluate the impact of this cause of death on this population. From 1997 to 2011, asphyxiation due to choking was identified as the cause of death in 14 of 350 cases (4%). Sampling of an unrelated but adjacent population over this same period yielded 186 necropsy cases of bottlenose dolphins with no cases of asphyxiation. Asphyxiated animals presented with a fish lodged in the cranial esophagus associated with a dislocated and obstructed or compressed larynx. There was no clear sex predilection. Affected animals included 12 adults and two juveniles. The fish species involved included sheepshead, black chin tilapia and striped mojarra. In five cases, recreational fishing gear was also present. Cetacean choking is related to selection of prey fish species with strong dorsal spines and may be secondarily associated with fish attached to fishing gear. Prey abundance and dolphin behavior may influence these selections. Environmental alterations leading to changes in prey availability or increased interactions with fishing gear may change the significance of fatal choking in dolphin populations.     

Overfishing of small pelagic fishes increases trophic overlap between immature and mature striped dolphins in the Mediterranean Sea

The interactions among diet, ecology, physiology, and biochemistry affect N and C stable isotope signatures in animal tissues. Here, we examined if ecological segregation among animals in relation to sex and age existed by analyzing the signatures of δ(15)N and δ(13)C in the muscle of Western Mediterranean striped dolphins. Moreover, we used a Bayesian mixing model to study diet composition and investigated potential dietary changes over the last two decades in this population. For this, we compared isotope signatures in samples of stranded dolphins obtained during two epizootic events occurring in 1990 and 2007-2008. Mean δ(13)C values for females and males were not significantly different, but age-related variation indicated δ(13)C enrichment in both sexes, suggesting that females and males most likely fed in the same general areas, increasing their consumption of benthic prey with age. Enrichment of δ(15)N was only observed in females, suggesting a preference for larger or higher trophic level prey than males, which could reflect different nutritional requirements. δ(13)C values showed no temporal variation, although the mean δ(15)N signature decreased from 1990 to 2007-2008, which could indicate a dietary shift in the striped dolphin over the last two decades. The results of SIAR indicated that in 1990, hake and sardine together contributed to 60% on the diet of immature striped dolphins, and close to 90% for mature striped dolphins. Conversely, the diet of both groups in 2007-2008 was more diverse, as hake and sardine contributed to less than 40% of the entire diet. These results suggest a dietary change that was possibly related to changes in food availability, which is consistent with the depletion of sardine stocks by fishing.     

Mass stranding of striped dolphin, Stenella coeruleoalba, at Augusta, Western Australia: notes on clinical pathology and general observations.

Seventeen striped dolphins, Stenella coeruleoalba, were found stranded on a West Australian beach. Three animals died before a rescue attempt was made and a further three died during the rescue. The remaining dolphins were released 24 km offshore and were not seen again. One dolphin was noted to have a broken mandible. Evidence of physical trauma to the other dolphins was minimal; one adult female was observed with some peeling skin. Blood was collected for analysis. All dolphins were slightly dehydrated and had a leukogram typical of a stressed animal. Plasma biochemistry reflected primary muscle trauma. There were no clues to the cause of the stranding; observed pathology reflected damage that occurred as a direct consequence of stranding.     

Post-epizootic chronic dolphin morbillivirus infection in Mediterranean striped dolphins Stenella coeruleoalba

Dolphin morbillivirus (DMV) has caused 2 epizootics with high mortality rates on the Spanish Mediterranean coast, in 1990 and 2006-07, mainly affecting striped dolphins Stenella coeruleoalba. Following the first epizootic unusual DMV infections affecting only the central nervous system of striped dolphins were found, with histological features similar to subacute sclerosing panencephalitis and old dog encephalitis, the chronic latent localised infections caused by defective forms of measles virus and canine distemper virus, respectively. Between 2008 and 2010, monitoring by microscopic and immunohistochemical (IHC) studies of 118 striped dolphins stranded along Catalonia, the Valencia Region and Andalusia showed similar localised DMV nervous system infections in 25.0, 28.6 and 27.4% of cases, respectively, with no significant differences among regions or sex. The body length of DMV-infected dolphins was statistically greater than that of non-infected dolphins (196.5 vs. 160.5 cm; p < 0.001). Molecular detection of DMV was performed by 2 different RT-PCR techniques amplifying a 429 bp fragment and a 78 bp fragment both within the phosphoprotein (P) gene. The 429 bp RT-PCR results contradicted the IHC-DMV results as only 3 of 6 dolphins with positive IHC-DMV had positive PCR results. All 6 cases were positive with the 78 bp RT-PCR. These findings contraindicate the use of the 429 bp RT-PCR protocol based on the P gene to detect this specific form of DMV. DMV localised nervous infection constitutes the most relevant single cause of stranding and death in Mediterranean striped dolphins in the years following a DMV epizootic, and it might even overwhelm the effects of the epizootic itself, at least in 2007.     

Evidence for infanticide in bottlenose dolphins of the western North Atlantic

Nine bottlenose dolphin (Tursiops truncatus) calves that stranded in Virginia in 1996 and 1997 died of severe blunt-force trauma. Injuries were concentrated on the head and chest and multiple rib fractures, lung lacerations, and soft tissue contusions were prominent. Skeletal and/or soft tissue trauma occurred bilaterally in all of the calves. One had a bite wound across the left mandible that exhibited deep punctures consistent with the tooth placement in an adult bottlenose dolphin. The lesions were not compatible with predation, boat strike, fisheries interactions, rough-surf injury, or blast injury. However, they were similar to traumatic injuries described in stranded bottlenose dolphin calves and harbor porpoises (Phocoena phocoena) in Great Britain attributed to violent dolphin interactions. The evidence suggests that violent dolphin behavior was the cause of the trauma in the nine calves reported here and that infanticide occurs in bottlenose dolphins of the western North Atlantic.     

一头搁浅瑞氏海豚的病理解剖和死因分析

瑞氏海豚在我国大陆沿海搁浅的案例相对较少。2019年7月10日,一头活体瑞氏海豚搁浅于广东省湛江市徐闻县石马角海域,半小时左右死亡。本文描述了该海豚的外部形态特征和骨骼系统,并通过解剖对其死因进行了分析。该瑞氏海豚为成年雌性,体长252.0 cm,体重约130.0 kg;体型消瘦,皮下脂肪层菲薄,体表无明显可致死外伤。头骨长45.6 cm,脊椎式为C7+T12+L23+Ca24=66,肋骨12对,V形骨19枚。解剖发现,该海豚的呼吸道和消化道中均存在大量泥沙;前胃内有两只长度超过30 cm的橡胶手套,胃和肠中几乎没有食糜和粪便。推测该瑞氏海豚可能因误食橡胶手套造成胃部阻塞而无法正常进食和消化,导致脂肪层过度消耗,体力不支而搁浅;在搁浅过程中可能因挣扎导致呼吸道进入大量泥沙引起呼吸不畅,最终死亡。当前难降解的海洋垃圾对海洋哺乳动物,包括以头足类为食的鲸类动物所产生的直接威胁已不容忽视。     

MITOCHONDRIAL DNA VARIABILITY OF STRIPED DOLPHINS (STENELLA COERULEOALBA) IN THE SPANISH MEDITERRANEAN WATERS

Frozen muscle samples from 44 striped dolphins stranded on the Spanish Mediterranean coasts from 1990 to 1993 have been studied by means of mitochondrial DNA (mtDNA) restriction site analysis. Thirty-five of these dolphins were affected by a die-off occurring during this time in the western Mediterranean Sea. The mtDNA from each dolphin was digested with 15 restriction endonucleases that recognized 61 different restriction sites. The specific location of these sites on the mitochondrial gene map allowed us to determine the distribution of variability along this molecule. From the restriction analysis, a total of 15 different composite patterns or haplotypes was obtained and their phylogenetic relationships established both by cladistic and phenetic methods.
Estimates of sequence divergence among the 15 haplotypes ranged from 0.148% to 0.919%. Haplotype diversity (average heterozygosity) obtained was 0.789, a high value compared to other available data on cetaceans, especially considering that only a small area of the distribution of the species has been sampled. Neither population subdivision nor any evidence of correlation between haplotype frequencies and temporal or geographic distribution of dolphins has been detected.     

Tetranucleotide microsatellite loci from the striped dolphin (Stenella coeruleoalba Meyen, 1833)

Five tetranucleotide microsatellite loci were isolated from the striped dolphin (Stenella coeruleoalba). Characterization of these loci revealed between three and 11 alleles at each locus and observed heterozygosity ranged from 0.192 to 0.775. All sets of primers amplified homologous loci when tested on four related species and showed polymorphisms in most locus–species combinations. A sixth microsatellite, which appeared to be monomorphic in the striped dolphin, was also shown to be polymorphic in three related species.     

The presence of mercury selenide in various tissues of the striped dolphin: evidence from μ-XRF-XRD and XAFS analyses

Marine mammals accumulate mercury in their tissues at high concentration and detoxify by forming mercury selenide (HgSe, tiemannite) mainly in the liver. We investigated the possibility of formation of HgSe in various tissues (liver, kidney, lung, spleen, pancreas, muscle and brain) other than the liver of the striped dolphin (Stenella coeruleoalba). We applied a combination method of micro-X-ray fluorescence (μ-XRF) imaging and micro-X-ray diffraction (μ-XRD) using a synchrotron radiation X-ray microbeam to analyze the tissue samples directly with minimal sample preparation. By this method, many accumulation points for Hg and Se on a micron scale were found in thin sections of the spleen and liver tissue and consequently, the XRF spectra and the XRD pattern of the hot spots confirmed the presence of tiemannite, HgSe. On the other hand, the insoluble fractions after enzyme digestion of the nuclear and mitochondrial fractions of all tissues were subjected to X-ray absorption fine structure (XAFS) analysis. XAFS analysis confirmed the presence of HgSe in all the tissues examined (liver, kidney, lung, spleen, pancreas, muscle and brain) of the striped dolphin. The presence of HgSe in all the tissues examined suggests that Se would be involved in the detoxification process of Hg in various tissues other than the liver. This contribution seems to be large especially in the liver and spleen but relatively small in the kidney, pancreas and brain, because the proportion of insoluble fraction containing HgSe was lower in these tissues (25 to 46%). This is the first report on the presence of tiemannite HgSe in various tissues of marine mammals.     

Characterization of alpha-fetoprotein in fetal striped dolphin (Stenella coeruleoalba): purification of protein product and molecular cloning of the corresponding transcript

Alpha-fetoprotein (AFP) is a fetal glycoprotein that is known as a biomarker for monitoring pregnancy in many mammalian species. However, characterization of AFP has not yet been undertaken in any cetacean species. Here, we purified AFP from the serum of fetal striped dolphin by chemical precipitation followed by a combination of immunoadsorbent column chromatography and gel filtration. The molecular masses of native and denatured dolphin AFP were estimated to be ～78,000 Da by gel filtration and ～68,000 Da by SDS-PAGE, respectively, representing typical masses reported for mammalian AFPs. In fetal serum, only the AFP band (～68,000 Da) appeared to be immunoreactive to an antiserum against purified dolphin AFP, indicating sufficient specificity for the development of an AFP immunoassay. Full-length cDNA encoding for the dolphin AFP was cloned from fetal liver and revealed an open reading frame comprising 610 amino acid residues, which included a putative signal peptide of 18 amino acid residues. This was followed by a sequence identical to the N-terminus of purified AFP. The deduced amino acid sequence of dolphin AFP showed more than 80% identity to those of other mammalian AFPs. To our knowledge, the present report represents the first identification and characterization of AFP from any cetacean species.     

Striped dolphins and short‐beaked common dolphins in the Gulf of Corinth,Greece: Abundance estimates from dorsal fin photographs

Capture‐recapture methods relying on dorsal fin natural markings have never been applied successfully to striped dolphins, Stenella coeruleoalba, and were rarely used to assess abundance of short‐beaked common dolphins, Delphinus delphis. We used digital photo‐identification to obtain abundance estimates of striped and common dolphins living in mixed groups in the Gulf of Corinth, Greece. The proportion of either species was calculated based on the relative number of photographs of adult animals showing relevant portions of their body during conspicuous surfacings. Striped dolphins and common dolphins averaged 95.0% and 3.2% of all individuals, respectively. Animals showing intermediate pigmentation accounted for another 1.8%. Striped dolphin numbers were relatively high, with a point estimate of 835 animals (95% CI = 631–1,106). Common dolphins numbers were low (point estimate 28 animals; 95% CI = 11–73) and individuals were scattered within striped dolphin groups, indicating that this common dolphin population may be nonviable. Within a semiclosed Gulf exposed to considerable anthropogenic impact, the future of both dolphin species is of concern due to their suspected geographic isolation and restricted extent of occurrence. Information provided here can be used to inform timely conservation efforts.