Abstract
Leptospirosis is the most widespread zoonotic disease in the world. It is caused by pathogenic spirochetes of the genus Leptospira spp. and is maintained in nature through chronic renal infection of carrier animals. Rodents and other small mammals are the main reservoirs. Information on leptospirosis in marine mammals is scarce; however, cases of leptospirosis have been documented in pinniped populations from the Pacific coast of North America from southern California to British Columbia. We report the isolation of a Leptospira spp. strain, here named Manara, from a kidney sample obtained from a Southern Right Whale (Eubalaena australis) calf, which stranded dead in Playa Manara, Península Valdés, Argentina. This strain showed motility and morphology typical of the genus Leptospira spp. under dark-field microscopy; and grew in Ellinghausen-McCullough-Johnson-Harris (EMJH) medium and Fletcher medium after 90 days of incubation at 28°C. Considering the source of this bacterium, we tested its ability to grow in Fletcher medium diluted with seawater at different percentages (1%, 3%, 5%, 7% and 10% v/v). Bacterial growth was detected 48 h after inoculation of Fletcher medium supplemented with 5% sea water, demonstrating the halophilic nature of the strain Manara. Phylogenetic analysis of 16S rRNA gene sequences placed this novel strain within the radiation of the pathogenic species of the genus Leptospira spp., with sequence similarities within the range 97–100%, and closely related to L. interrogans. Two different PCR protocols targeting genus-specific pathogenic genes (G1-G2, B64I-B64II and LigB) gave positive results, which indicates that the strain Manara is likely pathogenic. Further studies are needed to confirm this possibility as well as determine its serogroup. These results could modify our understanding of the epidemiology of this zoonosis. Until now, the resistance and ability to grow in seawater for long periods of time had been proven for the strain Muggia of L. biflexa, a saprophytic species. To the best of our knowledge, this is the first isolation of a Leptospira sp. from cetaceans. Our phenotypic data indicate that strain Manara represents a novel species of the genus Leptospira, for which the name Leptospira brihuegai sp. nov. is proposed.
Highlights
Leptospirosis is a neglected zoonotic disease, endemic in most tropical and subtropical regions of the world
The causative agents of this zoonosis are pathogenic strains belonging to the order Spirochaetales, family Leptospiracea and genus Leptospira spp
Seropositivity has been reported in North America, in California sea lions (Zalophus californianus) [4], Northern fur seals (Callorhinus ursinus) [5], Northern elephant seals (Mirounga angustirostiris) [6], and harbor seals (Phoca vitulina) [7,8]
Summary
Leptospirosis is a neglected zoonotic disease, endemic in most tropical and subtropical regions of the world. Leptospirosis is maintained in nature through chronic renal infection of carrier animals, with rodents and other small mammals as the most important reservoirs [1,2,3]. Leptospira spp. can cause disease in pinnipeds [9,10,11,12], periodic large scale stranding and mortality events, every three to four years, of California sea lions along the pacific coast of North America from southern California to British Columbia have been attributed to L. interrogans serovar Pomona infections [4, 10, 11, 12, 13,14]. Seroprevalence in marine mammals has been reported in the Pacific coast of South America, in Peruvian Amazon manatees (Trichechus inunguis) [15] and in Chilean South American sea lions (Otaria byronia) [16]
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