Distemper in a Dolphin

To the Editor: Deaths caused by new members of the genus Morbillivirus, family Paramyxoviridae (1), have occurred in recent decades among phocine and cetacean species, particularly harbor seals (Phoca vitulina) in 1988 (2) and 2002 (3). Endangered Mediterranean striped dolphins (Stenella coeruleoalba) died in 1990 and 1991 (4), and common dolphins (Delphinus delphis ponticus) from the Black Sea died in 1994 because of infection with dolphin morbillivirus (DMV) (5). A similar virus caused deaths in bottlenose dolphins (Tursiops truncatus) in the Gulf of Mexico from 1987 through 1994 (6). Closely related morbilliviruses caused deaths in harbor porpoises (Phocoena phocoena) in European waters in 1988 (7) (Porpoise morbillivirus) and endangered Mediterranean monk seals (Monachus monachus) in 1997 (8) (Monk seal morbillivirus). After these epidemics, the viruses disappeared and no marine or terrestrial reservoirs have been identified.


Distemper in a Dolphin
To the Editor: Deaths caused by new members of the genus Morbillivirus, family Paramyxoviridae (1), have occurred in recent decades among phocine and cetacean species, particularly harbor seals (Phoca vitulina) in 1988 (2) and 2002 (3).Endangered Mediterranean striped dolphins (Stenella coeruleoalba) died in 1990 and 1991 (4), and common dolphins (Delphinus delphis ponticus) from the Black Sea died in 1994 because of infection with dolphin morbillivirus (DMV) (5).A similar virus caused deaths in bottlenose dolphins (Tursiops truncatus) in the Gulf of Mexico from 1987 through 1994 (6).Closely related morbilliviruses caused deaths in harbor porpoises (Phocoena phocoena) in European waters in 1988 (7) (Porpoise morbillivirus) and endangered Mediterranean monk seals (Monachus monachus) in 1997 (8) (Monk seal morbillivirus).After these epidemics, the viruses disappeared and no marine or terrestrial reservoirs have been identifi ed.
In January 2007, a moribund, subadult, white-beaked dolphin (Lagenorhynchus albirostris) was found stranded on the North Friesian coast of Germany.The animal was humanely killed and a complete necropsy was performed.The main lesion was a nonsuppurative meningoencephalitis with neuronal degeneration and few eosinophilic cytoplasmic inclusion bodies characteristic of a viral disease.Lungs showed suppurative and interstitial pneumonia.Paraffi n-embedded sections of brain were examined for morbillivirus antigen by using an immunoperoxidase technique.We used various monoclonal antibodies that recognize different morbilliviruses.Tissues from a seal infected with phocine distemper virus and a dog with canine distemper were used as positive controls.Tissues from a white-beaked dolphin that underwent an autopsy in 2006 were used as negative controls.In the diseased dolphin, morbillivirus antigen was found exclusively in neurons and glial cells of the brain (Figure , panel A).
Frozen tissue samples and blood were examined for morbillivirus nucleic acid by reverse transcription-PCR with a set of universal morbillivirus primers that are specifi c for highly conserved regions of virus nucleoprotein (N) (9) and phosphoprotein (P) (10).A 457-bp amplicon of the P gene (GenBank accession no.EF451565) and a 287-bp amplicon of the N gene (GenBank accession no.EF469546) were detected in brain tissue.Our isolate, DMV/DE/2007, showed homologies of 99% with the N gene and 98% with the P gene of DMV isolated from Mediterranean striped dolphins.Phylogenetic analysis showed that isolate DMV/DE/2007 is closely related to DMV (Figure, panel B), porpoise morbillivirus, and monk seal morbillivirus (8).
Histologic changes in the dolphin resembled those of distemper in seals (3), porpoises (7), and other dolphins (4)(5)(6).Identifi cation of morbillivirus antigen in diseased tissues and isolation of genome fragments of a morbillivirus provide conclusive evidence for a primary etiologic role of this virus.Sequencing of the virus and phy-logenetic comparison showed that the virus is closely related to previously described dolphin morbillivirus and porpoise and monk seal morbilliviruses (8).To our knowledge, this is the fi rst report of morbillivirus infection in a white-beaked dolphin in German waters and in a marine mammal since the last epidemic among harbor seals in northern Europe in 2002.Isolation of DMV has not been reported since 1994.
Our fi ndings indicate that DMV is still circulating in some marine mammals.Similar to infections in terrestrial hosts, morbillivirus infections may occur in marine mammals in cycles without overt clinical disease in susceptible animals, as documented for harbor seals (2,3).Serum samples collected from 1995 through 1999 from cetacean species in various regions were positive for DMV, but porpoises and striped dolphins showed a decrease in humoral immunity, making them vulnerable to new epidemics.No data exist on seroprevalence of morbillivirus-specifi c antibodies  in white-beaked dolphins.We do not know how the dolphin contracted the infection and whether this remains an isolated case or the beginning of a new zoonosis.
White-beaked dolphins are found in moderate and subarctic waters of the Atlantic Ocean between the eastern coast of North America and northern Europe.They may migrate hundreds of kilometers within days.Therefore, these dolphins may play a role as a reservoir and vector for this morbillivirus, which is infectious for harbor porpoises, bottlenose dolphins, and other cetacean species (10).The reappearance of a morbillivirus represents a serious threat to susceptible marine mammals in northern European and American waters, with potentially devastating consequences and possibly the beginning of a new epidemic.

Bartonella australis sp. nov. from Kangaroos, Australia
To the Editor: During April-May 1999, 3 Bartonella isolates (AUST/ NH1, AUST/NH2, AUST/NH3) were cultivated and established from the blood of 5 Macropus giganteus gray kangaroos from central coastal Queensland, Australia.We used multigene sequencing to evaluate whether these Bartonella isolates fulfi ll the minimum requirements for classifi cation as a new species.
DNA from each Bartonella isolate was extracted by using the QIAamp tissue kit (QIAGEN, Hilden, Germany) according to the manufacturer's instructions.Partial PCR amplifi cation and sequencing of the genes encoding the 16S rDNA (rrs), citrate synthase (gltA), β-subunit of the RNA polymerase (rpoB), and cell division protein (ftsZ), as well as for the 16S-23S rDNA intergenic spacer (ITS) were attempted by using previously described primers and conditions (1).Bartonella sp.isolates AUST/NH1 to AUST/NH3 exhibited identical sequences for all 4 genes and the spacer studied, and isolate AUST/NH1 was selected as type strain among kangaroo isolates.Similarity rates between strain Aust/NH1 and validated Bartonella species (online Appendix Table, available from www.cdc.gov/EID/content/13/12/1961-appT.htm) ranged from 84.7% to 91.6%, from 97.5% to 98.5%, from 79.6% to 87.2%, from 85.4% to 95.0%, and from 83.5% to 87.1% for the ITS and rrs, gltA, rpoB, and ftsZ genes, respectively.Therefore, for each of these 4 genes or the spacer, strain AUST/NH1 exhibited similarity rates with all other species lower than the cutoffs published to classify Bartonella isolates within a validated species (1).It may thus be regarded as a new species.
The phylogenetic relationships among the studied bartonellae were inferred from sequence alignments of each gene and from concatenated gene sequences by using the maximum parsimony and neighbor-joining methods within the MEGA version 2.1 software package (3) and the maximumlikelihood method within the PHYLIP software package (4).Using rrs, gltA, and rpoB sequences, the phylogenetic position of strain AUST/NH1 was supported by bootstrap values <70%.In contrast, by using the ITS, ftsZ, and concatenated sequences, strain

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Cost-effectiveness analysis of alternative human papillomavirus vaccination strategies* Letters reporting cases, outbreaks, or original research should contain no more than 800 words and 10 references.They may have one Figure or Table and should not be divided into sections.All letters should contain material not previously published and include a word count.