First record of Trichinella in Leopardus guigna (Carnivora, Felidae) and Galictis cuja (Carnivora, Mustelidae): new hosts in Chile.

BACKGROUND: Trichinellosis is a zoonotic disease with a worldwide distribution. It is caused by several species of nematodes in the genus Trichinella. Trichinella spp. are transmitted through predation or carrion consumption and occur in domestic and sylvatic cycles. In humans trichinellosis occurs due to the consumption of raw or undercooked, infected meat and is mainly associated with the household slaughter of pigs or the consumption of game animals without veterinary inspection, a cultural practice that is difficult to resolve. Therefore, knowledge of this parasite's reservoir is relevant for better implementing public health strategies. The aim of this study was to assess the presence of Trichinella sp. in several carnivore and omnivore vertebrates in central-southern Chile. METHODS: We collected muscle tissue from a total of 53 animals from 15 species and were digested to detect Trichinella larvae which were further identified to species level using molecular techniques. RESULTS: We detected Trichinella larvae in Leopardus guigna (Felidae) and Galictis cuja (Mustelidae). We identified the larvae collected from L. guigna as Trichinella spiralis, but we were unable to molecularly characterize the larvae from G. cuja. This is the first record of Trichinella in a native mustelid of South America and the first record of T. spiralis in L. guigna. This study identified two novel hosts; however, further work is needed to identify the role that these and other hosts play in the cycle of Trichinella in Chile.

Survey of Trichinella in American minks (Neovison vison Schreber, 1777) and wild rodents (Muridae and Cricetidae) in Chile.

Trichinellosis is a zoonosis with worldwide distribution. Several hosts have been recognized around the world; however, there is a lack of knowledge of the role of feral mammals in Chile in its transmission. Herein, we tested muscle samples from 555 individuals among American minks (Neovison vison Schreber, 1777. n = 100) and several myomorph rodent species (Muridae and Cricetidae. n = 455) from southern Chile by artificial digestion to detect Trichinella larvae and identified the larvae at the species level through molecular analyses. Rodents were captured in agricultural and wild protected areas of several administrative regions (hereafter: region), while minks were captured in agricultural areas of the Los Ríos region. Trichinella spiralis larvae were detected in a synanthropic black rat (0.24%) of the Ñuble region and in seven minks (7%) trapped in agricultural areas of the Los Ríos region. The present results suggest that T. spiralis circulation is restricted to synanthropic rodents and minks living in agricultural areas where the parasite circulates among domesticated pigs. This study represents the first record of T. spiralis in a mustelid mammal in South America, increasing the number of feral species that could participate in the reservoir.

Trichinella spiralis in a cougar (Puma concolor) hunted by poachers in Chile.

Trichinellosis is a zoonosis caused by nematodes of the Trichinella genus, of which 10 species have been described. Species identification when larvae is found is only possible using molecular tools. Meat from pigs and game animals not subjected to veterinary inspection are the main sources of human infections. The hunting of native carnivores is prohibited in Chile due to conservation issues and the fact that those animals favor pest control. The illegal hunting of a cougar (Puma concolor) occurred in September 2020. Herein, the molecular identification of Trichinella larvae, by analyzing nuclear (expansion segment V) and mitochondrial (cytochrome C oxidase subunit I) sequences are described. Both the amplification of the expansion segment V region and the phylogenetic analysis of a segment of a fragment of the cytochrome c-oxidase subunit I sequence confirmed that the larvae belonged to T. spiralis. The case described herein represents the first evidence of illegal hunting of a protected mammal infected with Trichinella in Chile, highlighting the 'One Health' perspective to face this disease in the rural-sylvatic interphase.

Trichinella spiralis in a cougar (Puma concolor) hunted by poachers in Chile / Trichinella spiralis em um puma (Puma concolor) caçado por caçadores furtivos no Chile

Abstract Trichinellosis is a zoonosis caused by nematodes of the Trichinella genus, of which 10 species have been described. Species identification when larvae is found is only possible using molecular tools. Meat from pigs and game animals not subjected to veterinary inspection are the main sources of human infections. The hunting of native carnivores is prohibited in Chile due to conservation issues and the fact that those animals favor pest control. The illegal hunting of a cougar (Puma concolor) occurred in September 2020. Herein, the molecular identification of Trichinella larvae, by analyzing nuclear (expansion segment V) and mitochondrial (cytochrome C oxidase subunit I) sequences are described. Both the amplification of the expansion segment V region and the phylogenetic analysis of a segment of a fragment of the cytochrome c-oxidase subunit I sequence confirmed that the larvae belonged to T. spiralis. The case described herein represents the first evidence of illegal hunting of a protected mammal infected with Trichinella in Chile, highlighting the 'One Health' perspective to face this disease in the rural-sylvatic interphase.

Resumo A triquinelose é uma doença causada por nematoides do gênero Trichinella. Dez espécies foram descritas. A identificação das espécies a partir das larvas só foi possível com ferramentas moleculares. O abate doméstico de suínos e o consumo de animais de caça, sem inspeção veterinária, são as principais fontes de infecção humana. A caça de carnívoros nativos é proibida no Chile por questões de conservação e pelo fato de que esses animais favorecem o controle de pragas. A caça ilegal de um puma (Puma concolor) aconteceu em setembro de 2020. No Chile, a identificação molecular de larvas de Trichinella, obtida pelo exame de sequências nucleares (segmento de expansão V) e mitocondriais (citocromo C oxidase subunidade I) são descritas. Tanto a amplificação da região do segmento de expansão V quanto a análise filogenética de um segmento de um fragmento da sequência da subunidade I da Citocromo c-oxidase, confirmaram que as larvas pertenciam a T. spiralis. O caso aqui descrito representa a primeira evidência de caça ilegal de um mamífero protegido infectado com Trichinella no Chile, destacando a perspectiva de "Saúde Única" para enfrentar esta doença na interfase rural-silvestre.

Characterization of mesenchymal stem cells derived from adipose tissue of a cougar (Puma concolor).

Adipose derived mesenchymal stem cells (AMSCs) have been isolated from domestic and wild cats. For wild cats, the isolation of AMSCs has been reported in the black-footed cats (Felis nigripes) and guigna (Leopardus guigna). Stromal vascular fraction (SVF) isolated from cougar adipose tissue have been used to restore elbow functionality in the cougar (Puma concolor) but multipotent characteristics of these cells have not been described. The present study describes for the first time the isolation and characterization of mesenchymal stem cells derived from adipose tissue of cougar. AMSCs and fibroblasts from six months female cougar were isolated and cultured in DMEM/F12, supplemented with FBS 10% + 1% Antibiotic/Antifungal + 2.4 mM L-Glutamine + 2.4 mM pyruvate up to passage 5. Expression of pluripotent and surface marker genes was evaluated at mRNA level. Mesodermal differentiation (adipogenic, osteogenic and chondrogenic) was described. AMSCs expressed mRNA of pluripotent genes Oct4, Nanog, Sox2 and Klf4 and surface markers Cd44, Cd90, Cd105 and MHCII. Fibroblasts showed similar mRNA expression with the exception of Sox2. AMSCs obtained from cougar exhibit multipotency features similar to domestic cats MSC, nevertheless, other analyses are required. AMSCs from cougar could be a source of interest for treatment of individuals that remain in captivity or arrive to wildlife rehabilitation centers.