Search for antibodies against Trichinella in two synanthropic Procyonidae species from southeast Mexico: white-nosed coatis (Nasua narica) and raccoons (Procyon lotor).

Trichinella is a nematode that are spread by the consumption of parasitized meat. Carnivora, a mammalian order, serve as key hosts for this parasite. However, evidence of Trichinella in wildlife from the Neotropics is extremely scarce, with reports documenting its presence only for five carnivore species: two Felidae, one Otariidae and two Mustelidae. Other widely distributed species that are consumed as bushmeat, such as Procyonidae, have not been studied in this context. A long-term study was performed for antibodies against Trichinella in coatis (Nasua narica) and common raccoons (Procyon lotor) in southeastern Mexico. Between the summer of 2009 to the winter 2013, a total of 291 coati samples and 125 raccoon samples were collected from a tropical green area located within an urban zone. An Enzyme-linked immunosorbent assay (ELISA) was used to detect antibodies against the excretory and secretory products of Trichinella spiralis muscle larva. ELISA-positive samples were further confirmed by Western Blot analysis. Results showed no evidence of antibodies during the first two years of study. However, in 2011, a sudden appearance of anti-Trichinella occurred. The seroprevalence reached its highest peak of 43% for coatis during winter 2013 and 53% for raccoons in summer 2013. This is the first study that provides evidence of Trichinella circulation within a neotropical procyonid community.

Canine distemper in neotropical procyonids: Molecular evidence, humoral immune response and epidemiology.

Canine Distemper Virus (CDV) can produce a fatal multisystem disease in carnivores and other mammals and is an important threat for wildlife conservation. However, integrative and comparative studies in wild carnivores are scarce and some areas of the world lack of genetic studies. We explore the dynamic of host-CDV in a procyonid community during an outbreak. This study reports for the first time an index case occurred in a common raccoon (Procyon lotor) and for which a complete CDV diagnosis was performed. The long-term epidemiological analysis in two sympatric populations of common raccoons and white-nosed coatis (Nasua narica) was achieved through seroneutralization, RT-PCR and direct immunofluorescence assays. Additionally, hematologic analyses were performed and phylogenetic reconstruction of CDV was done using molecular data from this study. Overall prevalence for white-nosed coatis was 19.6 % and for common raccoons was 25.3 % by seroneutralization, and 13.3 % and 17.3 % by RT-PCR. Antibodies titer average for white-nosed coatis was 1:512 and 1:156 for common raccoons. Significant difference in prevalence between white-nosed coatis and common raccoons was detected during one season (summer 2013). White-nosed coatis showed differences in erythrocytes and monocytes counts between positives and negative animals. A 100 % similarity was found between CDV of white-nosed coati and CDV of common raccoon and is a new CDV sequence not previously described; this sequence is close to Asian and European lineage. An endemic state of distemper in both species was observed but showed different dynamics over time per host species. Differences in cellular and humoral responses were also detected between procyonids. The evidence found here may have serious implications for CDV understanding in wild carnivores, it reveals clear differences in the response over time to the same CDV strain, in two close related carnivore species.

Prevalence and Epitope Recognition of Anti-Trypanosoma cruzi Antibodies in Two Procyonid Species: Implications for Host Resistance.

More than 180 mammalian species have been found naturally infected with Trypanosoma cruzi. Many of them play an important role in the maintenance of this parasite. In particular, new studies have appeared which indicate that some species of Procyonidae family may play a role as T. cruzi hosts, however, more data are needed to evaluate their long-term physiological response to parasite infection, especially for specific antibodies. In this study, antibodies to T. cruzi were detected and prevalence and epitope recognition were assessed by ELISA (using discrete typing unit (DTU) I as antigen) and WB (using DTU I and DTU II as antigens) and sera from two procyonid species obtained through five-year follow-up of two semicaptive populations living in the same habitat. Marked heterogeneity in antigens recognition between species and differences in seroprevalence (p = 0.0002) between white-nosed coatis (Nasua narica), 51.8% (115/222), and common raccoons (Procyon lotor), 28.3% (23/81), were found. Antigens with high molecular weight when DTU-I was used were the most recognized, while a greater antigen diversity recognition was observed with DTU-II; for white-nosed coatis, low-molecular-weight antigens were mainly recognized, while for common raccoons proteins with molecular weights greater than 80 kDa were recognized most. These divergent humoral immune responses could be related to an alleged pattern of recognition receptors and major histocompatibility complex molecules difference in the procyonids species.

RADIOGRAPHIC ANATOMY OF THE FORELIMB IN THE NORTHERN TAMANDUA (TAMANDUA MEXICANA).

The thoracic limb anatomy of anteaters in the family Myrmecophagidae is specialized for accessing termite and ant nests and for defense purposes. In the case of the northern tamandua (Tamandua mexicana), the forelimbs are also adapted for arboreal and terrestrial locomotion. Unfortunately, this species faces many conservation threats, such as habitat loss and traffic accidents, and injured individuals are frequently taken to wildlife rehabilitation centers. However, lack of knowledge of the radiographic osteoanatomy of this species may prevent appropriate management of injuries and thereby reduce the chances of successful release and survival. In order to fill this knowledge gap, this article describes for the first time the radiographic anatomy of the thoracic limb of the northern tamandua using four standard views and one additional view. The additional orthogonal view helps visualize structures, such as the hamatus process and the sesamoid bone, that are otherwise difficult to visualize due to the natural forearm position of anteaters. Additionally, some fractures and physeal growth plates were identified in one juvenile individual. Further radiographic investigations should be conducted on anteaters to provide more tools for diagnosis, treatment, and rehabilitation of these animals.

Molecular genotyping of Blastocystis spp. in wild mammals from Mexico.

Blastocystis spp. are common intestinal parasites found worldwide in humans and a wide range of animals. They exhibit extensive genetic diversity; currently, 17 subtypes (STs) and some groups called non-mammalian and avian STs (NMASTs) have been proposed. In addition, a large variety of animals have been reported as hosts of the parasite, and new hosts and STs are still being described. In this study, Blastocystis infection of wild animals in two sylvatic areas of Mexico was surveyed. Of one hundred twenty-four fecal samples, six were positive for Blastocystis: specifically, one sample from an opossum, one sample from a bat, and four samples from different species of rodents. ST4, ST17, and nucleotide sequences similar to Blastocystis lapemi were identified based on SSU rDNA sequences. To our knowledge, this is the first report to investigate species poorly or not previously evaluated for Blastocystis infection. Mammals having different niches and geographical distribution were infected with similar genetic type of Blastocystis, so that we suggest that local water or food sources could play an important role in Blastocystis transmission and ST maintenance in wild animals. Additionally, there are STs with scarce genetic variation, suggesting that they could be highly adapted to their hosts. These data contribute to our understanding of the host range and genetic diversity of Blastocystis.

Comparative leucocyte populations between two sympatric carnivores (Nasua narica and Procyon lotor).

Coatis (Nasua narica) and raccoons (Procyon lotor) potentially play an important role in zoonotic diseases because they may carry pathogens and can transmit them to humans. To date, our understanding of the immune function of these two carnivores is deficient. The aim of this study was to compare the number of leucocyte subtypes and the phagocytic capacity between the coati and the raccoon. Blood samples were collected, and leucocyte subtypes were characterized and counted by flow cytometry and microscopy, respectively. Phagocytosis was analysed by kinetic assay. Differences in leucocytes between these two species were found; the total count of neutrophils was higher in raccoons than in coatis, but lymphocytes and eosinophils were higher in coatis than in raccoons. Antigen reduction was more rapid for the coatis. However, raccoons had a higher efficient endocytic process than coatis. This study provides the basis for understanding the procyonid immune system, which informs conservation, particularly since some procyonids are imperilled.

Epidemiological study of ticks collected from the northern tamandua (Tamandua mexicana) and a literature review of ticks of Myrmecophagidae anteaters.

The family Myrmecophagidae contains three anteater species: Tamandua mexicana (Saussure, 1860), Tamandua tetradactyla (Linnaeus, 1758) and Myrmecophaga tridactyla (Linnaeus, 1758). These American anteater species currently face many conservation threats, among which road traffic accidents stand out. Parasitic studies on this family are scarce, and some of them include records of ectoparasites. Specifically for northern tamandua (T. mexicana), there is a lack of studies at population level. The objectives of the present research were to carry out an epidemiological study of tick species and its abundance on road-killed northern anteater specimens and, moreover, to perform a literature review of ticks collected from anteaters of Myrmecophagidae family. Five tick species were identified, including four Amblyomma spp. and Rhipicephalus sanguineus sensu lato, on 23 road-killed anteaters. Tick infestation prevalence was 43% (10/23), with a median tick infestation intensity of 3.5 per anteater (interquartile range 1-13.7). The bibliographic review highlighted the existence of twenty-nine ixodid species recorded on the three anteater species from 14 countries, mainly Brazil. The most common tick species on the Myrmecophagidae family are Amblyomma nodosum, A. calcaratum, A. cajennense sensu lato and A. auricularium. Some of these ixodids were also described as vectors of pathogens. Further studies are needed to evaluate the impact of ticks on anteater fitness, and to assess the role of these mammals as reservoirs of vector-borne diseases.

The role of sloths and anteaters as Leishmania spp. reservoirs: a review and a newly described natural infection of Leishmania mexicana in the northern anteater.

For years, mammals of the order Pilosa have been considered Leishmania reservoirs. But while most studies have focused on sloth species, anteaters have been overlooked, and in many Leishmania endemic countries like Mexico, no studies have been carried out. The aims of this work were to identify the presence of Leishmania spp. in tissue samples from road-killed northern tamanduas (Tamandua mexicana), using PCR amplification and sequencing of ITS1 DNA, and to discuss the role of Pilosa mammals as reservoirs of Leishmania based on available scientific records. This is the first study that identifies Leishmania in T. mexicana, from 1 of 16 individuals analyzed, so the estimated prevalence (CI 95%) of infection was 6.3% (0.3-27.2). Amplified sequence exhibited a 98.9% (727/735) similarity with L. mexicana, and phylogenetic analysis grouped the species in the L. mexicana-amazonensis cluster. The literature review revealed 241 cases of Leishmania spp. infection among 1219 Pilosa mammals evaluated, with prevalence between studies ranging from 3.5% in the brown-throated three-toed sloth (Bradypus variegatus) to 78% in the Hoffman's two-toed sloth (Choloepus hoffmanni). Current scientific information indicates that C. hoffmanni sloths are reservoirs of Leishmania, and further studies are needed in order to clarify if other Pilosa species play a role in Leishmania transmission.

New Entamoeba group in howler monkeys (Alouatta spp.) associated with parasites of reptiles.

Our knowledge of the parasite species present in wildlife hosts is incomplete. Protozoans such as amoebae of the genus Entamoeba infect a large variety of vertebrate species, including NHPs. However, traditionally, their identification has been accomplished through microscopic evaluation; therefore, amoeba species have not always been identified correctly. We searched for Entamoeba spp. using a fragment of the small subunit rDNA in free-ranging howler monkeys (Alouatta palliata and A. pigra) from southeast Mexico. One hundred fifty five samples were collected, with 46 from A. palliata and 109 from A. pigra and 8 of the total samples were positive. We detected a new clade of Entamoeba, which was separated from other described species but closer to E. insolita, as well as an unnamed sequence typically found in iguana species with low shared identity values (<90%). We designated this new clade as conditional lineage 8 (CL8) and we have shown that members of this group are not exclusive to reptiles.

Clarifying the Cryptic Host Specificity of Blastocystis spp. Isolates from Alouatta palliata and A. pigra Howler Monkeys.

Although the presence of cryptic host specificity has been documented in Blastocystis, differences in infection rates and high genetic polymorphism within and between populations of some subtypes (ST) have impeded the clarification of the generalist or specialist specificity of this parasite. We assessed the genetic variability and host specificity of Blastocystis spp. in wild howler monkeys from two rainforest areas in the southeastern region of Mexico. Fecal samples of 225 Alouatta palliata (59) and A. pigra (166) monkeys, belonging to 16 sylvatic sites, were analyzed for infection with Blastocystis ST using a region of the small subunit rDNA (SSUrDNA) gene as a marker. Phylogenetic and genetic diversity analyses were performed according to the geographic areas where the monkeys were found. Blastocystis ST2 was the most abundant (91.9%), followed by ST1 and ST8 with 4.6% and 3.5%, respectively; no association between Blastocystis ST and Alouatta species was observed. SSUrDNA sequences in GenBank from human and non-human primates (NHP) were used as ST references and included in population analyses. The haplotype network trees exhibited different distributions: ST1 showed a generalist profile since several haplotypes from different animals were homogeneously distributed with few mutational changes. For ST2, a major dispersion center grouped the Mexican samples, and high mutational differences were observed between NHP. Furthermore, nucleotide and haplotype diversity values, as well as migration and genetic differentiation indexes, showed contrasting values for ST1 and ST2. These data suggest that ST1 populations are only minimally differentiated, while ST2 populations in humans are highly differentiated from those of NHP. The host generalist and specialist specificities exhibited by ST1 and ST2 Blastocystis populations indicate distinct adaptation processes. Because ST1 exhibits a generalist profile, this haplotype can be considered a metapopulation; in contrast, ST2 exists as a set of local populations with preferences for either humans or NHP.

Possible Differences in the Effects of Trypanosoma cruzi on Blood Cells and Serum Protein of Two Wildlife Reservoirs.

A key step in the dynamics of vector-borne diseases is the role of seasonality. Trypanosoma cruzi is a protozoan that causes Chagas disease. Some wild mammals are considered natural hosts, yet not all mammals show the same response to infection. We explored the effect of T. cruzi on blood parameters in two mammal carnivores, coati (Nasua narica) and raccoon (Procyon lotor), that were naturally infected in summer and winter seasons. The study was carried out in the Zoological Park "Parque Museo de la Venta," in Southeastern Mexico. Blood samples were collected in summer and winter from 2010 to 2013. Parasite infection was assessed by PCR from whole blood, and a complete hemogram was determined by traditional manual methods. We found that both species had the same T. cruzi I lineage. For coatis, mean corpuscular volume, mean corpuscular hemoglobin, and monocytes were dependent of season, while eosinophils and plasma proteins were significantly different, but with no season effect. For raccoon, erythrocytes, mean corpuscular volume, mean corpuscular hemoglobin, and monocytes were dependent of season. These results and a previous study that indicated interspecific differences in parasitemia in both species suggest that raccoon is a better reservoir than coati. Such a different interspecific response implies that animals do not contribute equally to maintain T. cruzi parasites in the ecosystem. Such inequality differs according to season.

ENDEMIC ORTHOPOXVIRUS CIRCULATING IN PROCYONIDS IN MEXICO.

Limited serosurveillance studies suggested that orthopoxviruses (OPXV) are widespread in the US (e.g., Raccoonpox virus, Skunkpox virus, Volepox virus) and Brazil (Vaccinia virus); however, their animal reservoir(s) remain unconfirmed. Mexican mammal diversity includes several species related to those in which evidence for OPXV infections has been found (Oryzomys, Peromyscus, Microtus, and Procyonidae). The presence of these groups of mammals in Mexico and the evidence of their possible involvement in the maintenance of OPXV in nature suggest the same or similar OPXV are circulating in Mexico. We tested 201 sera from 129 procyonids via modified enzyme-linked immunosorbent assay (ELISA) and Western blot (WB) to estimate OPXV antibody prevalence in these animals. We detected a prevalence of 16.67% in Nasua narica (white-nosed coati), 35% in Procyon lotor (raccoon), and 30.4% in Bassariscus astutus (ring-tailed cat) when tested by either ELISA or WB. Western blot results presented protein bands consistent with the size of some OPXV immunodominant bands (14, 18, 32, 36, and 62 kDa). These results support the hypothesis that OPXV circulate in at least three genera of Procyonidae in Central and Southeast Mexico.

Follow up of natural infection with Trypanosoma cruzi in two mammals species, Nasua narica and Procyon lotor (Carnivora: Procyonidae): evidence of infection control?

BACKGROUND: A large variety of mammals act as natural reservoirs of Trypanosoma cruzi (the causal agent of Chagas disease) across the American continent. Related issues are infection and parasite burden in these reservoirs, and whether they are able to control T. cruzi infections. These parameters can indicate the real role of mammals as T. cruzi reservoirs and transmitters. Here, two species of mammals, white-nosed coati (Nasua narica) and raccoon (Procyon lotor), were examined for to determine: a) T. cruzi presence, and; b) their ability to control T. cruzi infection. METHODS: Multiple capture-recaptures of both species were carried out in semi-wild conditions in Villahermosa, Tabasco, Mexico, for 5 years. Two samplings per year (summer and winter) took place. Prevalence and pattern of T. cruzi infection were determined by PCR from both mammals' blood samples. RESULTS: Raccoon samples had a higher relative infection values (26.6%) compared to those of white-nosed coati (9.05%), being this difference significant in summer 2012 (P < 0.00001), summer (P = 0.03) and winter 2013 (P = 0.02). Capture and recapture data indicated three infection dynamics: 1) negative-positive-negative infection; 2) positive-negative-positive infection; and 3) positive at all sampling times. CONCLUSIONS: These results indicate that both coati and raccoons may be able to control T. cruzi infection. Thus, the role as efficient reservoirs could be questioned (at least for those times when mammals are able to tolerate the infection). However, while infected, they may also be able to approach human dwellings and play a role important in linking sylvatic and domestic cycles.

Cross transmission of gastrointestinal nematodes between captive neotropical felids and feral cats.

Pathogen cross transmission between wildlife and domestic animals represents an extinction risk for wildlife; however, reliable verification is difficult to perform, and in some cases, it is even considered unlikely to be conducted. The aim of this work was to identify cross transmission of helminths between feral cats and captive wild felids at a zoological park in southeastern Mexico. Feces were collected from jaguars (Panthera onca), cougars (Puma concolor), ocelots (Leopardus pardalis), margays (Leopardus wiedii), and jaguarundis (Puma yagouaroundi). A flotation technique and macroscopic sieving were performed on the feces. Additionally, as part of the noxious fauna control program of the park, feral cats were captured and euthanized. To perform parasitologic studies, helminths from these animals were recovered. Toxocara cati and Trichuris campanula were shared by jaguarundis and feral cats. Ancylostoma sp. was found in jaguar and ocelot and Ancylostoma tubaeforme in feral cats. Additionally, during this study, a couple of jaguarundis died with clinical signs of trichuriasis. This is the first report of T. campanula in jaguarundi. Because feral cats roam freely in the park, transmission could occur from these vertebrates to wild felids. This study shows the risk that parasites represent to wild felids; a similar situation could be found in free-living species, especially in fragmented habitats that favor contact with domestic animals.