Molecular based confirmation of puma meat sausages implicated in trichinellosis outbreaks in Argentina.

Trichinellosis is a zoonotic disease caused by Trichinella, with the main source of infection being the consumption of pork and pork-derived products. However, it can also be acquired from eating the meat from wild animals targeted for sport hunting. The objectives of this study were: 1) to develop and implement a molecular method for the identification of Sus scrofa (pig and wild boar) and Puma concolor (Puma) meat in sausages eaten raw, which were linked to trichinellosis outbreaks occurring in Córdoba, Buenos Aires and La Pampa provinces, Argentina, in 2010, 2021, and 2022, respectively; and 2) to identify the Trichinella species present in the food. Specific primers were designed for PCR amplification and nucleotide sequencing of a region of the mitochondrial cytochrome b gene from both host species. Samples from the mentioned outbreaks were analysed, and the molecular identification of Trichinella spp. larvae was also performed. Results of the species identification system revealed that sausages from Córdoba and Buenos Aires had a mixed composition of pork and puma meat, while those from La Pampa contained puma meat only. Trichinella spiralis was implicated in all three outbreaks. The species identification system developed and implemented in this study revealed two host species of Trichinella related to human cases, and alerts about the risk of zoonotic transmission to humans through infected puma meat.

Cylicospirura felineus in Puma (Herpailurus yagouaroundi) in southern Brazil - Macroscopic and microscopic aspects.

Parasites of the Cylicospirura genus are known to affect domestic and wild carnivores by causing the formation of nodules within the gastric mucosa and pyloric region of parasitized animals. C. felineus is a specimen of this genus that has been reported in Brazil in animal species such as Leopardus geoffroyi and Puma concolor. However, as far as the authors of the present study are aware, reports of C. felineus in jaguarundis (Puma yagouaroundi) do not yet exist. This wild feline species native to the Americas is found only in a few countries - and it is possibly extinct in others, such as the United States. In Brazil, the P. yagouaroundi species is classified as vulnerable by government environmental agencies. Consequently, identifying the diseases and their relevance to this species is of utmost importance for P. yagouaroundi's preservation. This study aims to describe the histopathological lesions found in jaguarundis parasitized by the genus Cylicospirura, and to identify the parasitic species observed in these animals. Eight specimens of P. yagouaroundi necropsied between 2010 and 2020 in the Laboratory of Veterinary Pathology of the Federal University of Paraná (UFPR) in Palotina, Brazil, were studied. Seven specimens were wildlife roadkills, and one was from the Municipal Zoo of Cascavel, Paraná, Brazil. Seven of the necropsied animals presented - in the greater curvature of the stomach, near the cardia - one or more parasitic nodules of 0.5 to 3.0 cm in diameter that contained several intralesional reddish filiform parasites. Histologically, these nodules were characterized as transmural parasitic granulomas surrounded by reaction with deposition of dense connective tissue. The parasitological evaluations by light and scanning electron microscopy identified morphological C. felineus' characteristics through the presence of unequal spicules in males, the position of the vulva, and location of the pedunculated papillae. Despite the gastric lesion caused by the parasite, the apparent low pathogenicity to the gastrointestinal system of the evaluated animals, massive infections or large nodules can lead to death - other members of the Spirocercidae family are known to cause organ rupture, esophageal obstructions, and mesenchymal neoplasms. Therefore, C. felineus should be included in the list of gastric parasites with pathogenic potential in the species P. yagouaroundi.

Absence of Trichinella spp. larvae in carcasses of road-killed wild animals in Paraná state, Brazil.

Trichinella spp. are zoonotic parasites that are widely distributed in warm-blooded carnivores and omnivores, including humans. Until the present moment, Brazil has been considered by World Animal Health Organization free from the domestic cycle of trichinellosis, whereas the parasite's sylvatic cycle has the status of infection in limited zones. However, neighboring countries such as Argentina have reports of parasite larvae in the wild fauna. The present study aimed to determine the occurrence of Trichinella spp. in road-killed wild animals in Paraná, Brazil. Biological samples from 71 wild animals-29 Didelphis albiventris, 11 Nasua nasua, ten Cerdocyon thous, seven Dasypus novemcinctus, six Leopardus guttulus, six Sphiggurus spinosus and two Puma concolor-collected from November 2016 to November 2021 were subjected to artificial digestion, following the methodology described in the REGULATION (EC) No. 2075/2005. No Trichinella spp. larvae were detected in the carcasses of the road-killed wild animals. However, considering the wide spectrum of possible reservoirs that could act as a link between the sylvatic and domestic cycles and considering the current Brazilian status of sylvatic trichinellosis in limited zones, frequent monitoring of wild fauna remains necessary.

Natural infection and molecular detection of Cytauxzoon felis in a free-ranging Puma concolor in the state of Goiás, Brazil / Infecção natural e detecção molecular de Cytauxzoon felis em Puma concolor de vida livre no estado de Goiás, Brasil

The puma (Puma concolor Linnaeus, 1771), the most widely distributed felid species in the Americas, can be found in all Brazilian biomes. Nevertheless, few studies have focused on hemoparasites in this species. Cytauxzoon felis, a hemoparasite that can infect domestic cats, has also been described in wild felids in Brazil. To the best of our knowledge, this study is the first to diagnose the natural infection and molecular detection of C. felis in a P. concolor in the state of Goiás. This animal presented non-regenerative anemia and inclusion suggestive of piroplasmids within red blood cells. The amplified 551 bp fragment of partial Piroplasmida 18S rRNA gene sequence was 100% identical to corresponding sequences of C. felis available in GenBank. No specific treatment for cytauxzoonosis was administered, and after rehabilitation, the animal was reintroduced into the wild. This finding provides some evidence that P. concolor may act as a natural host of the parasite. The epidemiology, vector and pathogenicity of this hemoparasite in wild and domestic cats in Brazil deserves further investigation.

O puma (Puma concolor Linnaeus, 1771) tem a maior distribuição entre os felídeos das Américas e é encontrado em todos os biomas do Brasil. No entanto, poucos estudos têm se concentrado nos hemoparasitos nesta espécie. Cytauxzoon felis, um hemoparasito que pode infectar gatos domésticos, também foi descrito em felídeos selvagens no Brasil. A saber, este estudo é o primeiro diagnóstico de infecção natural e detecção molecular de C. felis em um P. concolor do estado de Goiás. Este animal apresentou anemia arregenerativa e inclusão de piroplasmídeos nos glóbulos vermelhos. A amplificação do fragmento de 551 pb da sequência parcial do gene Piroplasmorida 18S rRNA foi 100% idêntica a sequências correspondentes de C. felis disponíveis no GenBank. Nenhum tratamento específico para citauxzoonose foi administrado e, após a reabilitação, o animal foi reintroduzido na natureza. Essa descoberta fornece algumas evidências de que P. concolor pode atuar como um hospedeiro natural do parasito. A epidemiologia, vetor e patogenicidade deste hemoparasito em gatos selvagens e domésticos no Brasil merecem uma investigação mais aprofundada.

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

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.(AU)

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.(AU)

Helminthfauna of road-killed cougars (Puma concolor) from the Northeastern Region of São Paulo State, Brazil

As the second-largest Neotropical carnivore, cougars (Puma concolor) are especially important for maintenance of the biodiversity and ecosystem health. Five wild adult cougars (Puma concolor), found roadkilled in highways in the Northeastern region of São Paulo, were evaluated in search for parasites. Ten species representing nine families were identified. The most prevalent helminths were Uncinaria bidens, Lagochilascaris major, Spirometra sp., and Oncicola canis, followed by Cylicospirura subaequalis, Toxascaris leonina, Taenia omissa, Echinococcus sp., Filaroides sp. and Oncicola oncicola. It is important to note that some helminths found in this study, such as L. major, Spirometra sp., O. oncicola, O. canis, Echinococcus sp., T. leonina, C. subaequalis, and Filaroides sp. are known to affect domestic carnivores, which may indicate interaction between wild and domestic hosts. This study represents a new host record for four of the species found in cougars, U. bidens, L. major, O. canis, and Filaroides sp., and new locality records for U. bidens, T. leonina, C. subaequalis, and Filaroides sp(AU)

Como o segundo maior carnívoro neotropical, as onças-pardas são muito importantes para a manutenção da biodiversidade e a saúde do ecossistema. Cinco pumas adultos de vida livre (Puma concolor), mortos em rodovias na região Nordeste de São Paulo, foram avaliados quanto a presença de parasitas. Dez espécies, representando nove famílias foram identificadas. Os helmintos mais prevalentes foram Uncinaria bidens, Lagochilascaris major, Spirometra sp., e Oncicola canis, seguidos por Cylicospirura subaequalis, Toxascaris leonina, Taenia omissa, Echinococcus sp., Filaroides sp. e Oncicola oncicola. É importante salientar que alguns dos helmintos encontrados neste estudo, tais como Lagochilascaris major, Spirometra sp., O. oncicola, Oncicola canis, Echinococcus sp., Toxascaris leonina, Cylicospirura subaequalis, e Filaroides sp., afetam sabidamente carnívoros domésticos, o que pode indicar a interação entre hospedeiros selvagens e domésticos. Este estudo representa novos registros de hospedeiro para quatro espécies de helmintos, sendo elas U. bidens, L. major, O. canis, and Filaroides sp., além de novo registro de localidade para U. bidens, T. leonina, C. subaequalis, e Filaroides sp(AU)

Parasites in road-killed wild felines from North of Paraná state, Brazil / Parasitas em felinos selvagens mortos em estradas do norte do estado do Paraná, Brasil

This study aimed to identify the intestinal parasites of road-killed wild felines in the North Central and North, Paraná state, southern Brazil. The animals were monitored by sampling previously established transects. The places where the felines were run over were mapped, the animals were identified, and the gastrointestinal tract was evaluated. The feces were submitted to coproparasitological techniques of spontaneous sedimentation, floating in hypersaturated NaCl solution and centrifugal floating in zinc sulfate. All the parasitic structures detected were photomicrographed. In the coproparasitological analyses were identified oocysts of Cystoisospora spp., eggs of Ancylostomatidae, and Capillaria spp.; eggs of Aelurostrongylus spp., Toxocara spp., Physaloptera spp., Taenia spp., and Spirometra spp.; Aelurostrongylus abstrusus larvae; and eggs and adults of Ancylostoma cati and Taenia spp. One of the cats was parasitized by a flea of Ctenocephalides felis felis. Based on these results, the animals analyzed in this study supplied important samples for the evaluation of parasitic diversity of North of Paraná and suggested that this region may have conditions that allow the maintenance of these parasites life cycles in the environment and among wildlife.(AU)

O objetivo deste trabalho foi identificar os parasitas intestinais de felinos silvestres mortos em estradas nas regiões Norte Central e Norte, Paraná, sul do Brasil. Os animais foram monitorados por amostragem de transectos previamente estabelecidos. Os locais de atropelamento foram mapeados, os animais foram identificados e enviados para autópsias, durante as quais amostras de fezes foram coletadas e submetidas a técnicas coproparasitológicas de sedimentação espontânea, flutuação em solução hipersaturada de NaCl e flutuação por centrífugação em sulfato de zinco e fotografadas, quando estruturas parasitárias estavam presentes. Nas análises coproparasitológicas, foram identificados oocistos de Cystoisospora spp., ovos de Ancylostomatidae e Capillaria spp, Aelurostrongylus spp., Toxocara spp., Physaloptera spp., Taenia spp. e Spirometra spp.; larvas de Aelurostrongylus abstrusus; e ovos e adultos de Ancylostoma cati e Taenia spp. Um dos felídeos estava parasitado por Ctenocephalides felis felis. Com base nesses resultados, os animais analisados neste estudo forneceram amostras importantes para a avaliação da diversidade parasitária do Norte do Paraná e sugeriram que esta região pode apresentar condições que possibilitem a manutenção dos ciclos de vida desses parasitas no ambiente e entre a vida silvestre.(AU)

Intestinal parasites in free-living Puma concolor

Background: Studies on intestinal parasites in cougars Puma concolor are scarce, and most of the available information on the species has been derived from individuals in captivity. One of the greatest threats to the survival of wild cats is habitat loss caused by urban sprawl, agricultural matrix, and linear developments such as highways and railways, which expose animals to direct contact with humans and to a high risk of death. Given the lack of scientific data the objective of this study was to report on the occurrence of intestinal parasites in a cougar (P. concolor) specimen from the northwestern region of Paraná State, Brazil. Case: The carcass of a free-living cougar (P. concolor) individual was sent to the Interdisciplinary Science Museum (Museu Interdisciplinar de Ciências - MIC) of Paranaense University (Universidade Paranaense - UNIPAR); an individual was killed during a collision with an unidentified vehicle on highway PR-486, in the municipality of Mariluz (PR, Brazil). The geographical coordinates of the location where the animal was found are: 23°59′29″S, 53°8′47″W. This region is characterized by semi-deciduous seasonal forest remnants. After being identified on site and collected by the inspectors of the Paraná Environmental Institute (Instituto Ambiental do Paraná - IAP), the cadaver was donated to the MIC (by the IAP) for a necropsy to elucidate the cause of death. In this context, the cadaver was sent to the Animal Pathology section of the Department of Veterinary Medicine at UNIPAR. The animal was necropsied by performing the standard necropsy technique for carnivores: opening the intestine through the mesentery and exposing the mucosa from the duodenum to the colon...(AU)

Nodular and sclerosing gastritis caused by Cylicospirura felineus in a puma (Puma concolor).

An adult male puma (Puma concolor), hit by a car in an urban area, died three days later despite the therapeutic support provided. At necropsy, multiple firm nodules were identified in the gastric mucosa. The nodules were coated by an intact mucosa with a central opening from which reddish and cylindrical nematodes protruded into the lumen. Twenty-seven nematodes were retrieved for morphological and morphometric evaluations. During histopathological examination of the gastric tissue, the adult nematodes appear in longitudinal and transverse sections, surrounded by thick bands of collagen, interspersed with mixed inflammatory infiltrates. The nematodes had an eosinophilic cuticle with caudal serrated projections (bulbar type), coelomyarian musculature, pseudocoelom, and females with uterus containing numerous larvated eggs, characteristics consistent with the Cylicospirura genus. Morphologically, female nematodes had six large tricuspid teeth in the oral cavity and the vulva had an opening anterior to the esophagus-intestinal junction. Male nematodes had five pairs of small papillae near the tip of the tail. These findings were consistent with Cylicospirura felineus. This parasite should be included in the differential diagnosis of nodular gastric wall lesions in wild felids.

Oncicola venezuelensis (Marteau, 1977) (Acanthocephala: Oligacanthorhynchidae) in Puma concolor in Rio de Janeiro, Brazil.

Specimens of Oncicola venezuelensis (Marteau, 1977) were recovered from fragments of intestinal tissue of a female Puma concolar (Linn, 1771) found dead in Petrópolis, Rio de Janeiro in 2017. A total of 140 helminths were recovered. Five males and 5 females of the helminths were analyzed morphologically as well as 50 parasite eggs recovered in intestinal contents. Morphologically, these helminths were compatible with the genus Oncicola, because of the size and shape of the proboscis, the size and disposition of the lemnisci and the morphometry of the eggs, in which the external membrane of the shell was delicate and clear. From histopathology, the helminths were deeply embeded in the mucosa reaching up to the muscle layer. One specimen was also identified molecularly with universal primers that amplified the eukaryote region ITS1-5.8S-ITS2. The helminth showed 99% identity with the gene sequence of O. venezuelensis deposited in GenBank. It is important to emphasize, this parasite has been very little reported in the literature, which reinforces the importance of this report.

Intestinal parasites in free-living Puma concolor

Background: Studies on intestinal parasites in cougars Puma concolor are scarce, and most of the available information on the species has been derived from individuals in captivity. One of the greatest threats to the survival of wild cats is habitat loss caused by urban sprawl, agricultural matrix, and linear developments such as highways and railways, which expose animals to direct contact with humans and to a high risk of death. Given the lack of scientific data the objective of this study was to report on the occurrence of intestinal parasites in a cougar (P. concolor) specimen from the northwestern region of Paraná State, Brazil. Case: The carcass of a free-living cougar (P. concolor) individual was sent to the Interdisciplinary Science Museum (Museu Interdisciplinar de Ciências - MIC) of Paranaense University (Universidade Paranaense - UNIPAR); an individual was killed during a collision with an unidentified vehicle on highway PR-486, in the municipality of Mariluz (PR, Brazil). The geographical coordinates of the location where the animal was found are: 23°59′29″S, 53°8′47″W. This region is characterized by semi-deciduous seasonal forest remnants. After being identified on site and collected by the inspectors of the Paraná Environmental Institute (Instituto Ambiental do Paraná - IAP), the cadaver was donated to the MIC (by the IAP) for a necropsy to elucidate the cause of death. In this context, the cadaver was sent to the Animal Pathology section of the Department of Veterinary Medicine at UNIPAR. The animal was necropsied by performing the standard necropsy technique for carnivores: opening the intestine through the mesentery and exposing the mucosa from the duodenum to the colon...

Nodular and sclerosing gastritis caused by Cylicospirura felineus in a puma (Puma concolor) / Gastrite nodular esclerosante causada por Cylicospirura felineus em onça parda (Puma concolor)

An adult male puma (Puma concolor), hit by a car in an urban area, died three days later despite the therapeutic support provided. At necropsy, multiple firm nodules were identified in the gastric mucosa. The nodules were coated by an intact mucosa with a central opening from which reddish and cylindrical nematodes protruded into the lumen. Twenty-seven nematodes were retrieved for morphological and morphometric evaluations. During histopathological examination of the gastric tissue, the adult nematodes appear in longitudinal and transverse sections, surrounded by thick bands of collagen, interspersed with mixed inflammatory infiltrates. The nematodes had an eosinophilic cuticle with caudal serrated projections (bulbar type), coelomyarian musculature, pseudocoelom, and females with uterus containing numerous larvated eggs, characteristics consistent with the Cylicospirura genus. Morphologically, female nematodes had six large tricuspid teeth in the oral cavity and the vulva had an opening anterior to the esophagus–intestinal junction. Male nematodes had five pairs of small papillae near the tip of the tail. These findings were consistent with Cylicospirura felineus. This parasite should be included in the differential diagnosis of nodular gastric wall lesions in wild felids.(AU)

Uma onça parda (Puma concolor) foi encontrada em uma área urbana após atropelamento e, apesar do suporte terapêutico fornecido, o animal morreu três dias depois. No exame post-mortem, múltiplos nódulos firmes foram identificados na mucosa gástrica. Os nódulos eram revestidos por mucosa intacta com um orifício central, do qual se insinuavam nematódeos cilíndricos e avermelhados. Vinte e sete nematódeos foram recuperados para avaliação morfológica e morfométrica. Na avaliação histopatológica do tecido gástrico, os nematódeos adultos apareceram em cortes longitudinais e transversais, circundados por bandas espessas de colágeno, intercaladas por infiltrado inflamatório misto. Os nematódeos eram constituídos por cutícula eosinofílica, com projeções serrilhadas voltadas caudalmente (do tipo bulbar), musculatura celomiariana, pseudoceloma e, nas fêmeas, útero com numerosos ovos larvados cujos achados foram sugestivos do gênero Cylicospirura. Morfologicamente, a cavidade bucal continha seis grandes dentes trífidos, na fêmea, a abertura da vulva era anterior à junção esôfago intestinal; e os machos tinham cinco pares de pequenas papilas próximas à ponta da cauda. Esses achados foram consistentes com Cylicospirura felineus. Este parasita deve ser incluído no diagnóstico diferencial de lesões nodulares da parede gástrica em felinos selvagens.(AU)

Nodular and sclerosing gastritis caused by Cylicospirura felineus in a puma (Puma concolor) / Gastrite nodular esclerosante causada por Cylicospirura felineus em onça parda (Puma concolor)

Abstract An adult male puma (Puma concolor), hit by a car in an urban area, died three days later despite the therapeutic support provided. At necropsy, multiple firm nodules were identified in the gastric mucosa. The nodules were coated by an intact mucosa with a central opening from which reddish and cylindrical nematodes protruded into the lumen. Twenty-seven nematodes were retrieved for morphological and morphometric evaluations. During histopathological examination of the gastric tissue, the adult nematodes appear in longitudinal and transverse sections, surrounded by thick bands of collagen, interspersed with mixed inflammatory infiltrates. The nematodes had an eosinophilic cuticle with caudal serrated projections (bulbar type), coelomyarian musculature, pseudocoelom, and females with uterus containing numerous larvated eggs, characteristics consistent with the Cylicospirura genus. Morphologically, female nematodes had six large tricuspid teeth in the oral cavity and the vulva had an opening anterior to the esophagus-intestinal junction. Male nematodes had five pairs of small papillae near the tip of the tail. These findings were consistent with Cylicospirura felineus. This parasite should be included in the differential diagnosis of nodular gastric wall lesions in wild felids.

Resumo Uma onça parda (Puma concolor) foi encontrada em uma área urbana após atropelamento e, apesar do suporte terapêutico fornecido, o animal morreu três dias depois. No exame post-mortem, múltiplos nódulos firmes foram identificados na mucosa gástrica. Os nódulos eram revestidos por mucosa intacta com um orifício central, do qual se insinuavam nematódeos cilíndricos e avermelhados. Vinte e sete nematódeos foram recuperados para avaliação morfológica e morfométrica. Na avaliação histopatológica do tecido gástrico, os nematódeos adultos apareceram em cortes longitudinais e transversais, circundados por bandas espessas de colágeno, intercaladas por infiltrado inflamatório misto. Os nematódeos eram constituídos por cutícula eosinofílica, com projeções serrilhadas voltadas caudalmente (do tipo bulbar), musculatura celomiariana, pseudoceloma e, nas fêmeas, útero com numerosos ovos larvados cujos achados foram sugestivos do gênero Cylicospirura. Morfologicamente, a cavidade bucal continha seis grandes dentes trífidos, na fêmea, a abertura da vulva era anterior à junção esôfago intestinal; e os machos tinham cinco pares de pequenas papilas próximas à ponta da cauda. Esses achados foram consistentes com Cylicospirura felineus. Este parasita deve ser incluído no diagnóstico diferencial de lesões nodulares da parede gástrica em felinos selvagens.

Oncicola venezuelensis () (Acanthocephala: Oligacanthorhynchidae) in Puma concolor in Rio de Janeiro, Brazil / Oncicola venezuelensis () (Acanthocephala: Oligacanthorhynchidae) em Puma concolor no Rio de Janeiro, Brasil

Abstract Specimens of Oncicola venezuelensis (Marteau, 1977) were recovered from fragments of intestinal tissue of a female Puma concolar (Linn, 1771) found dead in Petrópolis, Rio de Janeiro in 2017. A total of 140 helminths were recovered. Five males and 5 females of the helminths were analyzed morphologically as well as 50 parasite eggs recovered in intestinal contents. Morphologically, these helminths were compatible with the genus Oncicola, because of the size and shape of the proboscis, the size and disposition of the lemnisci and the morphometry of the eggs, in which the external membrane of the shell was delicate and clear. From histopathology, the helminths were deeply embeded in the mucosa reaching up to the muscle layer. One specimen was also identified molecularly with universal primers that amplified the eukaryote region ITS1-5.8S-ITS2. The helminth showed 99% identity with the gene sequence of O. venezuelensis deposited in GenBank. It is important to emphasize, this parasite has been very little reported in the literature, which reinforces the importance of this report.

Resumo Espécimes de Oncicola venezuelensis (Marteau, 1997) foram recuperados de fragmentos do tecido intestinal de uma fêmea de Puma concolor (Linn, 1771) encontrada morta em Petrópolis, Rio de Janeiro, em 2017. Um total de 140 helmintos foram recuperados. Cinco machos e 5 cinco fêmeas dos helmintos foram analisados morfologicamente, bem como 50 ovos dos parasitos recuperados no conteúdo intestinal. Morfologicamente, esses helmintos eram compatíveis com o gênero Oncicola, devido ao tamanho e formato da probóscide, o tamanho e disposição do leminisco e a morfometria dos ovos, que apresentaram membrana externa da casca delicada e clara. A partir da histopatologia, pode-se verificar que os helmintos estavam profundamente inseridos na mucosa, atingindo até a camada muscular. Um espécime também foi identificado molecularmente com primers universais que amplificam a região ITS-1.5.8S.ITS-2. Após as análises moleculares, foi verificado que os helmintos apresentavam 99% de identidade com sequência gênica de O. venezuelensis que está depositada no Genbank. É importante enfatizar, que esse parasito foi muito pouco relatado na literatura, demonstrando a importância deste relato.

Ancient parasitic DNA reveals Toxascaris leonina presence in Final Pleistocene of South America.

Parasitological analysis of coprolites has allowed exploring ecological relationships in ancient times. Ancient DNA analysis contributes to the identification of coprolites and their parasites. Pleistocene mammalian carnivore coprolites were recovered from paleontological and archaeological site Peñas de las Trampas 1.1 in the southern Puna of Argentina. With the aim of exploring ancient ecological relationships, parasitological analysis was performed to one of them, dated to 16 573-17 002 calibrated years BP, with 95.4% probability. Parasite eggs attributed to Toxascaris sp. by morphological characters were isolated. DNA of coprolite and eggs was extracted to molecular identification. Ancient mitochondrial DNA analysis confirmed the zoological origin of the coprolite as Puma concolor and that of parasite eggs as Toxascaris leonina. This is the oldest molecular parasite record worldwide, and it supports the presence of this parasite since the Pleistocene in America. These findings have implications for the biogeographic history of parasites and for the natural history of the region.

Oslerus (Anafilaroides) sp. in a Jaguarundi (Puma yagouaroundi) from Brazil.

We report the occurrence of Oslerus (Anafilaroides) sp. parasitizing the lung of the jaguarundi (Puma yagouaroundi) in Brazil.

Trichinella spiralis in a South American sea lion (Otaria flavescens) from Patagonia, Argentina.

Trichinella spp. from a sylvatic cycle has been found in several animal species such as pumas (Puma concolor), armadillos (Chaetophractus villosus), rats (Rattus norvegicus), and wild boars (Sus scrofa) in Argentina. Moreover, Trichinella infection has been detected in a wide range of marine mammals around the world, including polar bears (Ursus maritimus) and walruses (Odobenus rosmarus). Until the present time, Trichinella spp. infection has not been detected in marine mammals of South America. Samples from four South American sea lions (Otaria flavescens) found dead in Rio Negro, Argentina, were analyzed by artificial digestion, and in the case of one animal, Trichinella larvae were identified at the species level by nested multiplex PCR as Trichinella spiralis. This is the first report of a Trichinella species infecting marine mammals from South America.

Apex Predators, Rockshelters, and Zoonoses in the Patagonian Holocene.

Carnivorous mammals are a trophic guild with an important role in the dissemination of parasite infective stages (larvae, eggs, cysts, and oocysts). In the present study, new samples of coprolites attributed to carnivorous mammals, obtained from 2 archaeological caves, were analyzed for the presence of parasites with the aim to increase the knowledge about parasites in rockshelters that could have spread to humans and other mammals. To this purpose, fragments of 3 coprolites from Cerro Casa de Piedra, cave 5 and cave 7, were examined. Coprolites were rehydrated in aqueous trisodium phosphate and processed by spontaneous sedimentation. High parasite richness was observed and new parasite species for archaeological contexts were found. The parasitological findings in Puma concolor coprolites associated with caves suggest the importance of these carnivores in the dissemination of parasites in areas with high re-use of space and steady conditions of temperature, humidity, and radiation.

Occurrence and phylogenetic analysis of Candidatus Mycoplasma haemominutum in wild felines from Paraná, Brazil / Ocorrência e análise filogenética de Candidatus Mycoplasma haemominutum em felinos selvagens no Paraná, Brasil

Hemoplasma infections are emerging and wild fauna can represent an important reservoir of these pathogens. However, there are very few epidemiological studies about the occurrence of hemoplasmas in wild cats around the world. The purpose of this study is twofold: (1) evaluate the occurrence and phylogeny of hemoplasmas in captive wild felines at a zoo in the state of Paraná, Brazil, and (2) verify the correlation between subpopulations of these bacteria and the hematological and biochemical parameters of the animals. PCR was used to detect hemoplasmas in the blood of three cougars (Puma concolor), a jaguar (Panthera onca), a tiger (Panthera tigris) and a lion (Panthera leo), followed by sequencing and phylogenetic analysis. The cougars and jaguar were found to be hemoplasma-positive by PCR. The phylogenetic analysis of the 16S rRNA gene sequences enabled the identification of genotypes of Candidatus Mycoplasma haemominutum circulating in this zoo. The identified sequences were closely related to hemoplasma sequences originating from domestic cats and other wild cats, but the infected cougars and jaguar were healthy and showed no hematological or biochemical changes. It was concluded that P. concolor and P. onca are exposed to Candidatus Mycoplasma haemominutum in Paraná, but further research is suggested to assess the resistance of wild cats to different hemoplasma subpopulations.

Infecções por hemoplasmas são emergentes e os animais selvagens podem representar um importante reservatório desses patógenos. Entretanto, são escassos os estudos epidemiológicos sobre a ocorrência de hemoplasmas em felinos selvagens em todo o mundo. O objetivo deste estudo foi (1) avaliar a ocorrência e a filogenia de hemoplasmas em felinos selvagens cativos em um zoológico do Estado do Paraná, Brasil, (2) e verificar a correlação entre as subpopulações destas bactérias e os parâmetros hematológicos e bioquímicos dos animais. A PCR foi utilizada para detecção de hemoplasmas em sangue de três pumas (Puma concolor), uma onça (Panthera onca), um tigre (Panthera tigris) e uma leoa (Panthera leo), seguida de sequenciamento e análise filogenética. Constatou-se que os pumas e a onça foram PCR-positivos para hemoplasma. A análise filogenética das sequências do gene 16S rRNA permitiu identificar genótipos de Candidatus Mycoplasma haemominutum circulando neste zoo. As sequências identificadas apresentaram relações estreitas com sequências de hemoplasmas procedentes de gatos domésticos e de outros felinos selvagens; contudo, os pumas e a onça infectados apresentaram-se saudáveis e sem alterações hematológicas ou bioquímicas. Conclui-se que P. concolor e P. onca são expostos à Candidatus Mycoplasma haemominutum no Paraná, porém sugere-se a realização de pesquisas futuras para avaliar a resistência dos felinos selvagens frente à diferentes subpopulações dehemoplasmas.

Occurrence and phylogenetic analysis of Candidatus Mycoplasma haemominutum in wild felines from Paraná, Brazil / Ocorrência e análise filogenética de Candidatus Mycoplasma haemominutum em felinos selvagens no Paraná, Brasil

Hemoplasma infections are emerging and wild fauna can represent an important reservoir of these pathogens. However, there are very few epidemiological studies about the occurrence of hemoplasmas in wild cats around the world. The purpose of this study is twofold: (1) evaluate the occurrence and phylogeny of hemoplasmas in captive wild felines at a zoo in the state of Paraná, Brazil, and (2) verify the correlation between subpopulations of these bacteria and the hematological and biochemical parameters of the animals. PCR was used to detect hemoplasmas in the blood of three cougars (Puma concolor), a jaguar (Panthera onca), a tiger (Panthera tigris) and a lion (Panthera leo), followed by sequencing and phylogenetic analysis. The cougars and jaguar were found to be hemoplasma-positive by PCR. The phylogenetic analysis of the 16S rRNA gene sequences enabled the identification of genotypes of Candidatus Mycoplasma haemominutum circulating in this zoo. The identified sequences were closely related to hemoplasma sequences originating from domestic cats and other wild cats, but the infected cougars and jaguar were healthy and showed no hematological or biochemical changes. It was concluded that P. concolor and P. onca are exposed to Candidatus Mycoplasma haemominutum in Paraná, but further research is suggested to assess the resistance of wild cats to different hemoplasma subpopulations.(AU)

Infecções por hemoplasmas são emergentes e os animais selvagens podem representar um importante reservatório desses patógenos. Entretanto, são escassos os estudos epidemiológicos sobre a ocorrência de hemoplasmas em felinos selvagens em todo o mundo. O objetivo deste estudo foi (1) avaliar a ocorrência e a filogenia de hemoplasmas em felinos selvagens cativos em um zoológico do Estado do Paraná, Brasil, (2) e verificar a correlação entre as subpopulações destas bactérias e os parâmetros hematológicos e bioquímicos dos animais. A PCR foi utilizada para detecção de hemoplasmas em sangue de três pumas (Puma concolor), uma onça (Panthera onca), um tigre (Panthera tigris) e uma leoa (Panthera leo), seguida de sequenciamento e análise filogenética. Constatou-se que os pumas e a onça foram PCR-positivos para hemoplasma. A análise filogenética das sequências do gene 16S rRNA permitiu identificar genótipos de Candidatus Mycoplasma haemominutum circulando neste zoo. As sequências identificadas apresentaram relações estreitas com sequências de hemoplasmas procedentes de gatos domésticos e de outros felinos selvagens; contudo, os pumas e a onça infectados apresentaram-se saudáveis e sem alterações hematológicas ou bioquímicas. Conclui-se que P. concolor e P. onca são expostos à Candidatus Mycoplasma haemominutum no Paraná, porém sugere-se a realização de pesquisas futuras para avaliar a resistência dos felinos selvagens frente à diferentes subpopulações dehemoplasmas.(AU)