Molecular Characterization of New Haplotype of Genus Sarcocystis in Seabirds from Magdalena Island, Southern Chile

SIMPLE SUMMARY: Sarcocystidae is a family of apicomplexan protozoa highly prevalent in vertebrates. The definitive hosts of sarcocystids eliminate oocysts or sporocysts that infect intermediate hosts. After infection, mature tissue cysts (sarcocysts) develop in intermediate hosts, mostly in muscle and neurological tissues. Sarcocysts are infectious for definitive hosts, which acquire them through carnivorous or scavenging habits. Intermediate hosts and definitive hosts are the natural hosts of sarcocystids in which infections are usually mildly or not symptomatic. In 2017, muscular and neurological tissues of 22 birds from Magdalena Islands, southern coast of Chile, were screened for the presence of DNA of sarcocystids. DNA of organisms of the genus Sarcocystis was identified in two Chilean skuas (Stercorarius chilensis). The genetic makeup of the parasite detected in skuas was unprecedented and probably represent a new species in the genus. It is well known that Sarcocystis may cause severe infections in aberrant hosts, which are susceptible animals that do not behave as natural hosts for the parasite and have low resistance to the infection, thus more studies are needed to characterize this parasitosis in skuas and other hosts to understand the epidemiology of the infection and its impact on the health of marine fauna. ABSTRACT: Evidence of sarcocystid infection was investigated in samples of 16 penguins (Spheniscus. magellanicus), four Dominican gulls (Larus dominicanus) and two Chilean skuas (Stercorarius chilensis) found in Madalenas Islands, Chile, in 2017. Samples of skeletal muscle, cardiac muscle and brain from all birds were screened by a pan-sarcocystid nested-PCR targeting a short fragment of the gene encoding the small ribosomal unit (nPCR-18Sa). The only two positive samples by nPCR-18Sa, both from skuas, were tested by a nested-PCR directed to the internal transcribed spacer 1 (nPCR-ITS1), also a pan-sarcocystidae nested-PCR, and to a nested-PCR directed to the B1 gene (nPCR-B1), for the exclusive detection of Toxoplasma gondii. The two nPCR-18Sa-positive samples were nPCR-ITS1-positive and nPCR-B1-negative. The nPCR-ITS1 nucleotide sequences from the two skuas, which were identical to each other, were revealed closely related to homologous sequences of Sarcocystis halieti, species found in seabirds of northern hemisphere. Larger fragments of genes encoding 18S and partial sequences of genes coding for cytochrome oxidase subunit 1 were also analyzed, corroborating ITS1 data. The haplotypes found in the skuas are unprecedent and closely related to species that use birds as the definitive host. Further studies need to be carried out to detect, identify and isolate this parasite to understand the epidemiology of the infection and its impact on the health of marine fauna.