A long-term study on free-ranging jaguar-tick interactions, featuring a novel report of Amblyomma incisum adult infestation.

Over nearly 12 years, we collected ticks from free-ranging jaguars (Panthera onca) and performed statistical analyses to comprehend the vector-host relationship throughout the seasons. We evaluated the presence and number of ticks, as well as their association with weight, age, and gender of captured jaguars in the Pantanal and Amazon biomes. Out of 100 captured jaguars (comprising 72 initial captures and 28 recaptures, with 41 females and 31 males), 77 were found to be infested by different tick species. We gathered a total of 1,002 ticks, categorized by the following species in descending order of abundance: Amblyomma sculptum, Amblyomma ovale, Rhipicephalus microplus, Amblyomma triste, Amblyomma cajennense sensu stricto, Amblyomma incisum, and Amblyomma spp. larvae. Apart from weight, statistical analysis indicated that age, gender and seasonality does not significantly affect the presence of different tick species in free ranging jaguars. Notably, A. sculptum adults were more abundant in the first semester, while A. sculptum nymphs and Amblyomma spp. larvae were mainly found during dry months, aligning with their expected life cycle stages. This is the first long-term study in jaguars to correlate seasonality and host factors and also the first time an adult of A. incisum is reported infesting a jaguar.

Mitochondrial Genome of Spirometra theileri Compared with Other Spirometra Species.

This study was carried out to provide information on the taxonomic classification and analysis of mitochondrial genomes of Spirometra theileri. One strobila of S. theileri was collected from the intestine of an African leopard (Panthera pardus) in the Maswa Game Reserve, Tanzania. The complete mtDNA sequence of S. theileri was 13,685 bp encoding 36 genes including 12 protein genes, 22 tRNAs and 2 rRNAs with absence of atp8. Divergences of 12 protein-coding genes were as follow: 14.9% between S. theileri and S. erinaceieuropaei, 14.7% between S. theileri and S. decipiens, and 14.5% between S. theileri with S. ranarum. Divergences of 12 proteins of S. theileri and S. erinaceieuropaei ranged from 2.3% in cox1 to 15.7% in nad5, while S. theileri varied from S. decipiens and S. ranarum by 1.3% in cox1 to 15.7% in nad3. Phylogenetic relationship of S. theileri with eucestodes inferred using the maximum likelihood and Bayesian inferences exhibited identical tree topologies. A clade composed of S. decipiens and S. ranarum formed a sister species to S. erinaceieuropaei, and S. theileri formed a sister species to all species in this clade. Within the diphyllobothridean clade, Dibothriocephalus, Diphyllobothrium and Spirometra formed a monophyletic group, and sister genera were well supported.

Furuncular Myiasis by Dermatobia hominis (Diptera: Oestridae) in Wild Jaguars in the Amazon Rainforest.

Two individuals of the jaguar, Panthera onca (L.), were captured near the municipality of Presidente Figueiredo, Brazilian Amazon, during the years of 2017 and 2018. The jaguars presented furuncular myiasis caused by the human botfly Dermatobia hominis (L.) on the rear thighs and tail. This is the first record of infestation of D. hominis in P. onca in the Amazon region.

Molecular characterisation and morphological description of two new species of Hepatozoon Miller, 1908 (Apicomplexa: Adeleorina: Hepatozoidae) infecting leukocytes of African leopards Panthera pardus pardus (L.).

BACKGROUND: The African leopard Panthera pardus pardus (L.) is currently listed as a vulnerable species on the IUCN (International Union for the Conservation of Nature) red list of threatened species due to ongoing population declines. This implies that leopard-specific parasites are also vulnerable to extinction. Intracellular apicomplexan haemoparasites from the genus Hepatozoon Miller, 1908 have been widely reported from wild carnivores in Africa, including non-specific reports from leopards. This paper describes two new haemogregarines in captive and wild leopards from South Africa and provides a tabular summary of these species in relation to species of Hepatozoon reported from mammalian carnivores. METHODS: Blood was collected from nine captive and eight wild leopards at various localities throughout South Africa. Thin blood smears were Giemsa-stained and screened for intraleukocytic haemoparasites. Gamont stages were micrographed and morphometrically compared with existing literature pertaining to infections in felid hosts. Haemogregarine specific primer set 4558F and 2733R was used to target the 18S rRNA gene for molecular analysis. Resulting sequences were compared to each other and with other available representative mammalian carnivore Hepatozoon sequences from GenBank. RESULTS: Two species of Hepatozoon were found in captive and wild leopards. Of the 17 leopards screened, eight were infected with one or both morphologically and genetically distinct haemogregarines. When compared with other species of Hepatozoon reported from felids, the two species from this study were morphometrically and molecularly distinct. Species of Hepatozoon from this study were observed to exclusively parasitize a particular type of leukocyte, with Hepatozoon luiperdjie n. sp. infecting neutrophils and Hepatozoon ingwe n. sp. infecting lymphocytes. Phylogenetic analysis showed that these haemogregarines are genetically distinct, with Hepatozoon luiperdjie n. sp. and Hepatozoon ingwe n. sp. falling in well supported separate clades. CONCLUSIONS: To our knowledge, this is the first morphometric and molecular description of Hepatozoon in captive and wild African leopards in South Africa. This study highlights the value of using both morphometric and molecular characteristics when describing species of Hepatozoon from felid hosts.

An initial coprological survey of parasitic fauna in the wild Amur leopard (Panthera pardus orientalis).

The Amur leopard, one of nine recently recognized subspecies of leopard, is still the most threatened by a stochastic procession of extinction. Evaluation of the potential danger to the conservation of the Amur leopard originating from disease urgently needs to be studied. Unfortunately, research on the potential risk to Amur leopards caused by disease is rare. In terms of parasitic diseases that affect this species, even basic data for parasitic fauna are absent. The aim of this study is to acquire this knowledge to improve the general understanding of Amur leopard parasites. Seven parasite species, including 3 nematodes (Toxocara cati, a capillarid-type parasite, and a Metastrongyloidea-type parasite), 2 cestodes (Spirometra sp. and Taenia sp.), 1 trematode (Paragonimus sp.), and 1 protozoan (Cystoisospora felis), were found in this research. Toxocara cati occurred most frequently, followed by Spirometra sp.

Severe infection caused by nymphs of Armillifer armillatus (Pentastomida, Porocephalidae) in a leopard, Panthera pardus, in the Kruger National Park, South Africa.

The necropsy of an adult male leopard, Panthera pardus, shot in the Kruger National Park, revealed the presence of large numbers of Armillifer armillatus nymphs in the intestine, liver, spleen, mesentery, peritoneal fold, mediastinum and lungs. The animal had been observed to be blind in the right eye and severely debilitated. The infection with A. armillatus clearly contributed to its emaciation and anaemia. Armillifer armillatus is a parasite of snakes, using mammals that form part of the snakes' prey as intermediate hosts. It is also one of the pentastomids with the highest zoonotic potential in Africa. It is unclear if the leopard's partial blindness and injuries of its extremities forced it to forego larger prey items for easier prey, such as snakes, and this in turn led to exposure to this unusual parasite, or if he had simply developed a preference for snakes. The incidental finding of A. armillatus in a large carnivore emphasises the importance of necropsies in expanding our knowledge on wildlife diseases.

Prevalence of Toxoplasma gondii parasite in captive Mexican jaguars determined by recombinant surface antigens (SAG1) and dense granular antigens (GRA1 and GRA7) in ELISA-based serodiagnosis.

Toxoplasma gondii is an obligate intracellular protozoan parasite that infects almost all warm-blooded animals, including humans, causing serious public health problems. In this study, the seroprevalence of T. gondii in captive jaguars in 10 Mexican zoos was determined using single and mixtures of recombinant surface antigens (SAG1) and dense granular antigens (GRA1 and GRA7) in immunoglobulin G (IgG) enzyme-linked immunosorbent assays (ELISAs). Their efficacy was compared with the tachyzoite lysate antigen. All recombinant antigens were characterised by high sensitivity (92.5-97.5%); the specificity of the IgG ELISAs was variable (83.3-91.6%). Mixtures of the two recombinant proteins were generally more reactive than single antigens. GRA7 + SAG1 showed the highest sensitivity (97.5%) and specificity (91.6%), almost perfect agreement (96.2%), and a kappa value of 0.89. An area under the curve value of 0.998 represented a highly accurate test with a cutoff value of 0.8. The seroprevalence of anti-T. gondii IgG antibodies in the single and mixed recombinant antigen ELISAs was 75.0-76.9%. This study shows that GRA7 + SAG1 can be successfully used to diagnose T. gondii infection in jaguars for effective monitoring of prevalence and for devising control methods and prevention strategies against toxoplasmosis.

First record of Lagochilascaris minor (Nematoda: Ascarididae) in Leopardus geoffroyi (Carnivora: Felidae) in Brazil / Primeiro registro de Lagochilascaris minor (Nematoda: Ascarididae) em Leopardus geoffroyi (Carnivora: Felidae) no Brasil

Abstract Lagochilascariasis is a parasitic disease caused by nematodes of the genus Lagochilascaris. These parasites occur in the neotropical region and their definitive hosts are wild animals, domestic dogs and felids and, accidentally, humans. Here, infection by Lagochilascaris minor is recorded for the first time in a wild felid, in Brazil.

Resumo A lagochilascaríase é uma doença parasitária causada por nematoides do gênero Lagochilascaris. Com distribuição neotropical, este parasito tem como hospedeiros definitivos animais silvestres, canídeos e felídeos domésticos e, acidentalmente, seres humanos. Registra-se a primeira ocorrência de Lagochilascaris minor em um felídeo silvestre no Brasil.

First record of Lagochilascaris minor (Nematoda: Ascarididae) in Leopardus geoffroyi (Carnivora: Felidae) in Brazil.

Lagochilascariasis is a parasitic disease caused by nematodes of the genus Lagochilascaris. These parasites occur in the neotropical region and their definitive hosts are wild animals, domestic dogs and felids and, accidentally, humans. Here, infection by Lagochilascaris minor is recorded for the first time in a wild felid, in Brazil.

Aminoacyl tRNA synthetases as potential drug targets of the Panthera pathogen Babesia.

BACKGROUND: A century ago, pantheras were abundant across Asia. Illegal hunting and trading along with loss of habitat have resulted in the designation of Panthera as a genus of endangered species. In addition to the onslaught from humans, pantheras are also susceptible to outbreaks of several infectious diseases, including babesiosis. The latter is a hemoprotozoan disease whose causative agents are the eukaryotic parasites of the apicomplexan genus Babesia. Babesiosis affects a varied range of animals including humans (Homo sapiens), bovines (e.g. Bos taurus), pantheras (e.g. Panthera tigris, P. leo, P. pardus) and equines. Babesia spp. are transmitted by the tick vector Ixodes scapularis or ticks of domestic animals, namely Rhipicephalus (Boophilus) microplus and R. (B.) decoloratus. At the level of protein translation within these organisms, the conserved aminoacyl tRNA synthetase (aaRS) family offers an opportunity to identify the sequence and structural differences in the host (Panthera) and parasites (Babesia spp.) in order to exploit these for drug targeting Babesia spp. METHODS: Using computational tools we investigated the genomes of Babesia spp. and Panthera tigris so as to annotate their aaRSs. The sequences were analysed and their subcellular localizations were predicted using Target P1.1, SignalP 3.0, TMHMM v.2.0 and Deeploc 1.0 web servers. Structure-based analysis of the aaRSs from P. tigris and its protozoan pathogens Babesia spp. was performed using Phyre2 and chimera. RESULTS: We identified 33 (B. bovis), 34 (B. microti), 33 (B. bigemina) and 33 (P. tigris) aaRSs in these respective organisms. Poor sequence identity (~ 20-50%) between aaRSs from Babesia spp. and P. tigris was observed and this merits future experiments to validate new drug targets against Babesia spp. CONCLUSIONS: Overall this work provides a foundation for experimental investigation of druggable aaRSs from Babesia sp. in an effort to control Babesiosis in Panthera.

A Clonorchiasis Case of a Leopard Cat, Prionailurus bengalensis euptilurus, Diagnosed by Ultrasonography and Egg Detection in Republic of Korea.

A clonorchiasis case in a captive leopard cat, Prionailurus bengalensis euptilurus, was confirmed by ultrasonographic findings and egg morphologies found in the bile juice sample in the Korea. The leopard cat was introduced from the wild habitat of Gyeongsangnam-do, to Cheongju Zoo in Cheongju-si, Chungcheongbuk-do, Korea in August 2014. Physical examinations were basically performed for quarantine and check-up health. The cat was comparatively good in health except anorexia. The cyst-like bile duct dilation and the increased echogenicity of gall bladder wall and hepatic parenchyma were observed by ultrasonography. Ultrasound-guided needle biopsy was conducted for collecting bile juice and the specimens were observed under light microscope. The numerous small trematode eggs were detected in the bile juice sample of the light microscopy. The eggs were 25-33 (28±3) µm by 18-22 (20±1) µm in size and showed typical characteristics of Clonorchis sinensis egg, i.e., a dominantly developed operculum, shoulder rim and dust-like wrinkles in surface. To treat the liver fluke infection, 20 mg/kg of praziquantel was orally administered only once to the case. Follow-up studies including fecal examinations were conducted during 2 years after treatment. But no more eggs were detected from the case. In the present study, we described the first clonorchiasis case of leopard cat, which was confirmed by ultrasonographic findings and egg morphologies from the bile juice sample in Korea.

Identity of Spirometra theileri from a Leopard (Panthera pardus) and Spotted Hyena (Crocuta crocuta) in Tanzania.

In the present study, a Spirometra species of Tanzania origin obtained from an African leopard (Panthera pardus) and spotted hyena (Crocuta crocuta) was identified based on molecular analysis of cytochrome c oxidase I (cox1) and NADH dehydrogenase subunit I (nad1) as well as by morphological observations of an adult tapeworm. One strobila and several segments of a Spirometra species were obtained from the intestine of an African male leopard (Panthera pardus) and spotted hyena (Crocuta Crocuta) in the Maswa Game Reserve of Tanzania. The morphological characteristics of S. theileri observed comprised 3 uterine loops on one side and 4 on the other side of the mid-line, a uterine pore situated posterior to the vagina and alternating irregularly either to the right or left of the latter, and vesicular seminis that were much smaller than other Spirometra species. Sequence differences in the cox1 and nad1 genes between S. theileri (Tanzania origin) and S. erinaceieuropaei were 10.1% (cox1) and 12.0% (nad1), while those of S. decipiens and S. ranarum were 9.6%, 9.8% (cox1) and 13.0%, 12.6% (nad1), respectively. The morphological features of the Tanzania-origin Spirometra specimens coincided with those of S. theileri, and the molecular data was also consistent with that of S. theileri, thereby demonstrating the distribution of S. theileri in Tanzania. This places the leopard (Panthera pardus) and spotted hyena (Crocuta Crocuta) as new definitive hosts of this spirometrid tapeworm.

Dioctophyme renale (Nematoda: Dioctophymatidae) in Leopardus geoffroyi (Carnivora: Felidae) in the Neotropical region / Dioctophyme renale (Nematoda: Dioctophymatidae) em Leopardus geoffroyi (Carnivora: Felidae) na região Neotropical

Abstract Parasitic diseases affecting wild carnivores remain largely unknown or poorly described. Dioctophymosis is a parasitosis caused by the nematode Dioctophyme renale that is found worldwide. It affects domestic and wild animals and has been reported frequently in Brazil. This paper reports dioctophymosis in a wild felid for the first time.

Resumo As doenças parasitárias que acometem os carnívoros silvestres ainda são desconhecidas ou pouco descritas. A dioctofimatose, parasitose de distribuição mundial causada pelo nematoide Dioctophyme renale, acomete animais domésticos e silvestres e tem sido relatada com frequência no Brasil. Este estudo reporta o primeiro caso de dioctofimatose em um felídeo silvestre.

Dioctophyme renale (Nematoda: Dioctophymatidae) in Leopardus geoffroyi (Carnivora: Felidae) in the Neotropical region.

Parasitic diseases affecting wild carnivores remain largely unknown or poorly described. Dioctophymosis is a parasitosis caused by the nematode Dioctophyme renale that is found worldwide. It affects domestic and wild animals and has been reported frequently in Brazil. This paper reports dioctophymosis in a wild felid for the first time.

Hepatozoon SPP. Infect Free-Ranging Jaguars (Panthera onca) in Brazil.

This study investigated the presence of Hepatozoon spp. in jaguars ( Panthera onca ) and domestic animals in the Cerrado, Amazon, and Pantanal biomes of Brazil. Between February 2000 and January 2010, blood samples were collected from 30 jaguars, 129 domestic dogs ( Canis lupus familiaris), and 22 domestic cats ( Felis catus ) for molecular tests. All of the jaguars from the Pantanal (n = 22) and Cerrado (n = 4) and 3 of 4 jaguars from the Amazon were positive for Hepatozoon spp. Domestic dogs (62.8%) and cats (31.8%) were also positive for the agent. Hepatozoon nucleotide sequences from jaguars and domestic cats grouped with other Hepatozoon felis, whereas Hepatozoon from domestic dogs showed high similarity to Hepatozoon canis. Different species of Amblyomma were identified as parasitizing the jaguars and may act as vectors for Hepatozoon spp. Jaguars from the 3 sites were healthy and did not seem to be threatened by the hemoparasite within its population or environments. Most likely, jaguars play an important role in the maintenance of Hepatozoon spp. in nature.

Seroprevalence of Toxoplasma gondii infection in zoo and domestic animals in Jiangxi Province, China.

Toxoplasma gondii is a zoonotic protozoan parasite that infects a wide range of warm-blooded animals throughout the world. In the present study, antibodies to T. gondii were determined using a commercial indirect hemagglutination (IHA) test in wild animals in a zoo. Three of 11 giraffes (Giraffa camelopardalis) (27%), 1 of 5 wolves (Canis lupus laniger) (20%), 1 of 6 hippopotamuses (Hippopotamus amphibious) (17%), and 2 of 9 tundra swans (Cygnus columbianus) (22%) were found to be positive. No antibodies were detected in leopards (Panthera pardus), wild geese (Anser cygnoides), and Eastern grey kangaroos (Macropus giganteus). Domestic species from 13 counties of Jiangxi Province, China were also investigated by an indirect hemagglutination (IHA) test. Thirty-five of 340 goats (10%), 94 of 560 water buffaloes (17%), and 4 of 35 cattle (11%) were found to be seropositive. This is the first report of T. gondii infection in animals kept in zoos and domestic animals in this province.

Serosurvey of Smooth Brucella, Leptospira spp. and Toxoplasma gondii in Free-Ranging Jaguars (Panthera onca) and Domestic Animals from Brazil.

This study investigated the exposure of jaguar populations and domestic animals to smooth Brucella, Leptospira spp. and Toxoplasma gondii in the Cerrado, Pantanal and Amazon biomes of Brazil. Between February 2000 and January 2010, serum samples from 31 jaguars (Panthera onca), 1,245 cattle (Bos taurus), 168 domestic dogs (Canis lupus familiaris) and 29 domestic cats (Felis catus) were collected and analysed by rose bengal test for smooth Brucella, microscopic agglutination test for Leptospira spp. and modified agglutination test for T. gondii. Cattle populations from all sites (9.88%) were exposed to smooth Brucella, but only one jaguar from Cerrado was exposed to this agent. Jaguars captured in the Cerrado (60.0%) and in the Pantanal (45.5%) were seropositive for different serovars of Leptospira spp., cattle (72.18%) and domestic dogs (13.1%) from the three sites and one domestic cat from Pantanal were also seropositive for the agent. The most prevalent serotype of Leptospira spp. identified in jaguars from the Cerrado (Grippotyphosa) and the Pantanal (Pomona) biomes were distinct from those found in the domestic animals sampled. Jaguars (100%), domestic dogs (38.28%) and domestic cats (82.76%) from the three areas were exposed to T. gondii. Our results show that brucellosis and leptospirosis could have been transmitted to jaguars by domestic animals; and jaguars probably play an important role in the maintenance of T. gondii in nature.

Exposure of free-living jaguars to Toxoplasma gondii, Neospora caninum and Sarcocystis neurona in the Brazilian Pantanal.

Toxoplasma gondii, Neospora caninum and Sarcocystis neurona are related apicomplexan parasites that cause reproductive and neurological disorders in a wide range of domestic and wild animals. In the present study, the immunofluorescence antibody test (IFAT) was used to investigate the presence of antibodies against T. gondii, N. caninum and S. neurona in the sera of 11 free-living jaguars (Panthera onca) in two protected areas in the Pantanal region of Mato Grosso state, Brazil. Ten jaguars (90.9%) showed seropositivity for T. gondii, eight (72.7%) for S. neurona, and seven (63.6%) for N. caninum antigens. Our findings reveal exposure of jaguars to these related coccidian parasites and circulation of these pathogens in this wild ecosystem. To the best of our knowledge, this is the first serological detection of N. caninum and S. neurona in free-living jaguars.

First record of intestinal parasites in a wild population of jaguar in the Brazilian Atlantic Forest.

Small and isolated wildlife populations may be more susceptible to disease, which makes illness an important issue to investigate regarding the conservation of large carnivores. Here, we present the results of the first investigation of intestinal parasites in one of the last remaining populations of jaguars in the Brazilian Atlantic Forest. We studied parasites from fecal samples using three different techniques for parasitological examination: floatation in saturated sodium chloride solution, sedimentation and formalin-ether centrifugation. Intestinal parasites were detected in 70% of the analyzed samples, and seven taxa (mean = 3.7 taxa/sample) were identified. All the groups of parasites that were identified have been recorded in previous jaguar studies. However, the records of Class Trematoda and nematodes Trichuridae are the first evidence of these groups of worms in free-ranging jaguars in Brazil. Although our results do not provide conclusive evidence on the health of this jaguar population, given its very small size (approximately 20 animals) we stress the need to properly understand the dynamics of disease in this wild population and to evaluate the risk of contracting new diseases from domestic species inhabiting the neighboring areas. These represent imperative actions for the successful conservation of this threatened population of jaguar.

First record of intestinal parasites in a wild population of jaguar in the Brazilian Atlantic Forest / Primeiros registros de parasitos intestinais em uma população silvestre de onça-pintada na Mata Atlântica Brasileira

Small and isolated wildlife populations may be more susceptible to disease, which makes illness an important issue to investigate regarding the conservation of large carnivores. Here, we present the results of the first investigation of intestinal parasites in one of the last remaining populations of jaguars in the Brazilian Atlantic Forest. We studied parasites from fecal samples using three different techniques for parasitological examination: floatation in saturated sodium chloride solution, sedimentation and formalin-ether centrifugation. Intestinal parasites were detected in 70% of the analyzed samples, and seven taxa (mean = 3.7 taxa/sample) were identified. All the groups of parasites that were identified have been recorded in previous jaguar studies. However, the records of Class Trematoda and nematodes Trichuridae are the first evidence of these groups of worms in free-ranging jaguars in Brazil. Although our results do not provide conclusive evidence on the health of this jaguar population, given its very small size (approximately 20 animals) we stress the need to properly understand the dynamics of disease in this wild population and to evaluate the risk of contracting new diseases from domestic species inhabiting the neighboring areas. These represent imperative actions for the successful conservation of this threatened population of jaguar.

Populações pequenas e isoladas são mais suscetíveis a doenças, o que torna a investigação de enfermidades uma questão importante para a conservação de grandes carnívoros. O presente estudo apresenta os resultados da primeira investigação de parasitos intestinais em uma das últimas populações remanescentes de onça-pintada na Mata Atlântica brasileira. Os parasitos foram obtidos a partir de amostras fecais, sendo utilizadas três técnicas para exame parasitológico: flutuação em solução saturada de cloreto de sódio (Método de Willis), sedimentação (Método de HPJ) e centrifugação com formol-éter. Parasitos intestinais foram detectados em 70% das amostras analisadas, tendo sido identificados sete táxons (média = 3,7 taxa/amostra). Todos os grupos de parasitos identificados foram mencionados em estudos anteriores realizados com onças-pintadas. Entretanto, os registros de Trematoda e nematoides Trichuridae aqui obtidos representam a primeira evidência desses grupos de vermes em onças-pintadas de vida livre no Brasil. Embora os resultados obtidos não sejam conclusivos quanto à saúde da população estudada, em virtude do tamanho populacional reduzido (aproximadamente 20 indivíduos), recomenda-se a investigação da dinâmica de doenças, bem como a avaliação do risco de contração de novas doenças a partir do contato com animais domésticos provenientes do entorno da área estudada. Essas investigações são fundamentais para a conservação dessa população ameaçada de onças-pintadas.