3D wildlife skull models for wildlife veterinary training.

Wildlife veterinarians are necessary for zoonotic diseases and species loss management, and there is a rising interest to enroll at veterinary schools with the wish to work in zoo and wildlife medicine. However, teaching wildlife is challenging due to the difficulty faced by universities to work with wild animal specimens. The aim of the present was to evaluate the understanding efficiency of some anatomical and behavioral aspects using 3D printed models of four wildlife species skulls, the kinkaju (Potos flavus), the white-nosed coati (Nasua narica), the northern anteater (Tamandua mexicana), and the nine-banded armadillo (Dasypus novemcinctus). This study was performed on 85 third-year veterinary students, divided into an experimental and a control group, who used and not used 3D printed skulls, respectively. Results show that the experimental group shows higher scores, in three of the four variables evaluated, than the control group. Then, 3D wildlife printed skulls constitute a promising teaching tool for veterinary students. In fact, it may be as good as real skulls, since new 3D printers can print on high endurance and firmness stock with high accuracy at reduced costs. In this context, it is important to encourage its use for the training of new generations and keep professionals up to date.

Novel Francisella-like endosymbiont and Anaplasma species from Amblyomma nodosum hosted by the anteater Tamandua Mexicana in Mexico.

The microbiome represents a complex network among the various members of the community of microorganisms that are associated with a host. The composition of the bacterial community is essential to supplement multiple metabolic pathways that the host lacks, particularly in organisms with blood-sucking habits such as ticks. On the other hand, some endosymbionts showed some competence with potentially pathogenic microorganisms. Francisella-like endosymbionts (FLEs) encompass a group of gamma-proteobacterias that are closely related to Francisella tularensis, but are usually apathogenic, which brings nutrients like vitamin B and other cofactors to the tick. It has been postulated that the main route of transmission of FLE is vertical; however, evidence has accumulated regarding the possible mechanism of horizontal transmission. Despite growing interest in knowledge of endosymbionts in the Neotropical region, the efforts related to the establishment of their inventory for tick communities are concentrated in South and Central America, with an important gap in knowledge in Mesoamerican countries such as Mexico. For this reason, the aim of this work was to evaluate the presence and diversity of endosymbionts in the highly host-specialized tick Amblyomma nodosum collected from the anteater Tamandua mexicana in Mexico. We analysed 36 A. nodosum for the presence of DNA of endosymbiont (Coxiella and Francisella) and pathogenic (Anaplasma, Borrelia, Ehrlichia and Rickettsia) bacteria. The presence of a member of the genus Francisella and Candidatus Anaplasma brasiliensis was demonstrated. Our findings provide information on the composition of A. nodosum's microbiome, increasing the inventory of bacterial species associated with this hard tick on the American continent.

The genus Rickettsia in Mexico: Current knowledge and perspectives.

The genus Rickettsia encompasses 35 valid species of intracellular, coccobacilli bacteria that can infect several eukaryotic taxa, causing multiple emerging and re-emerging diseases worldwide. This work aimed to gather and summarise the current knowledge about the genus Rickettsia in Mexico, updating the taxonomy of the bacteria and their hosts by including all the records available until 2020, to elucidate host-parasite relationships and determine the geographical distribution of each Rickettsia species present in the country. Until now, 14 species of Rickettsia belonging to four groups have been recorded in Mexico. These species have been associated with 26 arthropod species (14 hard ticks, three soft ticks, two sucking lice, and seven fleas) and 17 mammal species distributed over 30 states in Mexico. This work highlights the high biological inventory of rickettsias for Mexico and reinforces the need to approach the study of this group from a One Health perspective.

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.

Morphological and molecular characteristics of hemoparasites in vaillant's frogs (Lithobates vaillanti).

Amphibian populations are declining around the world, and the main reasons are the environmental changes and pathogens. However, there are few studies addressing the interaction and impact of the different pathogens that affect amphibians, such as hemoparasites. These parasites had been described as common in some amphibian species, but unfortunately, their description and characterization are unclear and scarcely spread. The objective of the present study was to evaluate the morphological and molecular characterization of hemoparasites present in vaillant's frogs. Seven frogs of Lithobates vaillanti were captured at the biological station La Florida in Tabasco, Mexico. Blood smears were performed, and results show that 100% of the animals have hemoparasites. Three types of hemoparasites were found. Eighty-five percent of the frogs were positive to Hepatozoon sp., 57% to Lankesterella sp., and 28% to Trypanosoma sp. According to the molecular analysis of the obtained sequences of Trypanosoma sp. and Hepatozoon sp., both protozoans were positioned in between the clusters of parasites of different geographical regions. Nevertheless, no species names were assigned to any of these parasites because more sequences and analysis are needed.

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.

Molecular detection of Rickettsia rickettsii in ticks associated with the bobcat (Lynx rufus) in northeast Mexico.

The study of rickettsial agents associated with ticks from wild felines is scarce. In Europe, three species of Rickettsia have been detected (Rickettsia helvetica, Rickettsia massiliae, and Rickettsia monacensis) in ticks collected from the Iberian lynx (Lynx pardinus). However, no studies have been conducted on another lynx species. For this reason, the aim of this study was to identify the diversity of Rickettsia species in ticks associated with bobcats (Lynx rufus) collected in the State of Tamaulipas, Mexico. During 1999 and 2004, nine bobcats from two municipalities of the state were trapped and visually inspected for the presence of ticks. A total of 95 ticks were collected from these lynxes. Ticks were preserved in 96% ethanol. Subsequently we identified the presence of Rickettsia DNA by the amplification of several fragments of genes 17 kDa, ompA and ompB. Recovered sequences were concatenated, aligned, and compared with those of reference deposited in GenBank. Additionally, a phylogenetic analysis was performed using the Maximum Likelihood method. The ticks were morphologically identified as belonging to the species Dermacentor variabilis. We selected a subset of 60 ticks which were examined, and 5% (3/60) were positive with an identity of 99% to sequences of R. rickettsii deposited in GenBank. The results obtained represent the first record of R. rickettsii in ticks associated with wild carnivores, and in particular with bobcats distributed in northeast of Mexico.

Rickettsia species in ticks that parasitize amphibians and reptiles: Novel report from Mexico and review of the worldwide record.

Ticks are obligate haematophagous ectoparasites that are associated with a wide range of vertebrate hosts, among them also reptiles and amphibians. They have dynamic ecological interactions with multiple microorganisms, ranging from endosymbionts to pathogens, such as the members of the genus Rickettsia. The aim of this work was to detect Rickettsia in ticks from amphibians and reptiles from southern Mexico by the amplification, sequencing and phylogenetic reconstruction of the gltA and ompA genes, and also to compile all the published records worldwide of Rickettsia associated with ticks attached to reptiles and amphibians, in order to elucidate the host-parasite relationships, and to identify the geographical distribution of each bacterial species. We record for the first time the presence of Rickettsia sp. strain Colombianensi and Rickettsia amblyommatis in ticks from several reptiles and amphibians collected in three new localities from the states of Guerrero and Veracruz, Mexico. Additionally, we here report 23 Rickettsia taxa associated with 18 tick species attached to 42 host taxa of amphibians and reptiles in 36 countries. Our findings increase the inventory of rickettsia reported in Mexico and summarizes the knowledge of these bacteria associated with ticks of this particular group of vertebrate host worldwide.

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 insights into the clinico-histopathological and molecular features of Pelecitus (Filarioidea: Onchocercidae) from a raptor bird.

Some Onchocercidae nematodes such as Pelecitus are parasites of medical and veterinary importance. The adult stage of Pelecitus has been reported infecting birds, and the microfilaria has been associated to human blindness. However, in some of these cases, the nematode was incompletely identified at the species level due to the scarcity of morphological taxonomic keys and, also, to the lack of molecular diagnostic analysis. Here, we report a new Pelecitus species in a crested caracara (Caracara cheriway) producing a severe tenosynovitis and microfilarial dermatitis. It is also the first record of Pelecitus in an American bird of prey. Clinical and histopathological features are described, contributing towards our understanding of the pathogenesis of Pelecitus and the health and conservation of wild bird populations. Our study also provides new information on the molecular diagnosis of this parasite and highlights the potential role of wild birds as Pelecitus reservoirs, and health risk for humans and wildlife.

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.

Effect of host species diversity on multiparasite systems in rodent communities.

Reduced species diversity has been suggested to increase transmission rates and prevalence of infectious diseases. While this theory has been studied mostly in single pathogen systems, little is known regarding multiple pathogens systems in vertebrates at the community level. The aim of this study was to evaluate the effect of host richness and diversity on multiple parasite systems on a local scale. We captured small rodents and collected feces in three different vegetation types in a natural protected area in Janos, Chihuahua, Mexico. The flotation technique was used to identify parasite eggs or oocysts. Analysis of linear correlations was conducted between parasite prevalence and host and parasite diversity and richness. Negative correlation was detected between parasite prevalence and host diversity (p = 0.02 r(2) =-0.86), but no significant correlations was detected between parasite prevalence and host richness or parasite diversity or richness. Our study shows that at local scale, host diversity could affect multiple parasite systems in the same way that single pathogens do. Further studies should be performed on larger temporal and spatial scales to more thoroughly investigate the correlation observed in our analysis.

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.