Selected Wildlife Trematodes.

The trematodes are a species-rich group of parasites, with some estimates suggesting that there are more than 24,000 species. However, the complexities associated with their taxonomic status and nomenclature can hinder explorations of the biology of wildlife trematodes, including fundamental aspects such as host use, life cycle variation, pathology, and disease. In this chapter, we review work on selected trematodes of amphibians, birds, mammals, and their snail intermediate hosts, with the goal of providing a tool kit on how to study trematodes of wildlife. We provide a brief introduction to each group of wildlife trematodes, followed by some examples of the challenges each group of trematodes has relative to the goal of their identification and understanding of the biology and interactions these organisms have with their wildlife hosts.

Into the danger zone: How the within-host distribution of parasites controls virulence.

Despite the importance of virulence in epidemiological theory, the relative contributions of host and parasite to virulence outcomes remain poorly understood. Here, we use reciprocal cross experiments to disentangle the influence of host and parasite on core virulence components-infection and pathology-and understand dramatic differences in parasite-induced malformations in California amphibians. Surveys across 319 populations revealed that amphibians' malformation risk was 2.7× greater in low-elevation ponds, even while controlling for trematode infection load. Factorial experiments revealed that parasites from low-elevation sites induced higher per-parasite pathology (reduced host survival and growth), whereas there were no effects of host source on resistance or tolerance. Parasite populations also exhibited marked differences in within-host distribution: ~90% of low-elevation cysts aggregated around the hind limbs, relative to <60% from high-elevation. This offers a novel, mechanistic basis for regional variation in parasite-induced malformations while promoting a framework for partitioning host and parasite contributions to virulence.

A New Species of Cosmocerca Diesing, 1861 (Nematoda: Cosmocercidae) in Elachistocleis haroi Pereyra, Akmentins, Laufer, and Vaira, 2013 (Amphibia: Microhylidae) from South American Chaco.

PURPOSE: Cosmocercids are common nematodes that parasitize the digestive tract and lungs of amphibians and reptiles around the world. They are commonly found in leptodactylid and bufonid anurans in South America, primarily in Brazil and Argentina. This paper describes a new species of genus Cosmocerca based on specimens collected in a microhylid from the Dry Chaco ecoregion. METHODS: A total of 18 specimens of Elachistocleis haroi were collected in October 2011 in Chaco province and in December 2021 and March 2022 in Formosa province, Argentina, both areas of the Dry Chaco ecoregion. The morphology of the nematodes was studied in detail using light microscopy and scanning electron microscopy. RESULTS: One hundred and fifty-one nematodes were collected from the large intestines of E. haroi. Based on the morphology of the caudal papillae (rosette papillae and plectanes), the presence of two spicules and the absence of caudal alae in males, and the presence of two prodelphic ovaries in females, these specimens were allocated to the genus Cosmocerca. However, they presented unique characteristics that differentiate them from all known species. Cosmocerca wichiorum sp. nov. is similar to C. archeyi, C. australis, C. sardiniae, C. zugi, and C. leytensis by having four pairs of plectanes, but can be easily distinguished from all of them by the type, number, and arrangement of the precloacal (non-plectanes), adcloacal and postcloacal papillae, by the length of the spicules and by its different zoogeographical distribution. CONCLUSION: This is the 14th species of the genus Cosmocerca described in South America and the first one for the microhylid E. haroi from the Dry Chaco ecoregion.

GENETIC DIVERSITY OF CYSTODISCUS SPECIES IN AMPHIBIANS IN THE SOUTHERN UNITED STATES.

Myxosporean species in the genus Cystodiscus are parasites of amphibians and have been reported from several continents. Typically used for the identification of myxozoans, the spores produced by these species are similar to one another, possessing 2 polar capsules and being ovoid. The number of transverse depressions on the spore can be useful for delineating species, but these can sometimes be difficult to distinguish. In North America, Cystodiscus serotinus and Cystodiscus melleni have been described, and for C. serotinus in particular, numerous reports and a wide range of hosts have been associated with this species. Given the challenges of identifying some of these species, we questioned whether all encounters of Cystodiscus species can be attributed to these 2 described species, or if there may be additional undescribed species or cryptic species. Over 7 yr, 383 amphibians representing 13 species of toads, frogs, and salamanders were collected from sites in Oklahoma and Arkansas. Cystodiscus infections were found in 56 individuals (14.6%). Tissues from these infected individuals were preserved in alcohol for genetic analysis. The small subunit (SSU) and large subunit (LSU) ribosomal RNA genes were partially sequenced and analyzed phylogenetically. Nine distinct SSU sequence types and 7 distinct LSU sequence types were identified. Phylogenetically, sequence types were attributable to C. serotinus, C. melleni, Cystodiscus axonis, and an undescribed species. For the previously described species, there were multiple SSU sequence types: 4 for C. serotinus and 2 for both C. melleni and C. axonis. Phylogenetic patterns were similar for the LSU sequence analysis using a shorter sequence than the SSU, and we propose that the LSU is useful for initial barcoding of Cystodiscus species in any future surveys. In our qualitative assessment of sequence types compared to geography and host species, SSU types C1 and C2 (C. axonis) were only found in Union County, Arkansas, and McCurtain County, Oklahoma, respectively. Also, salamanders were only infected with SSU types B or D (C. melleni), and type B was only found in salamanders. Our finding of C. axonis in North America is notable because this species was described in Australia and is associated with host pathology. Our work reveals that there are cryptic species of Cystodiscus in the United States, one of which may be a pathogen, highlighting the importance of genetic analysis for future surveys of these species.

Host-multiparasite interactions in amphibians: a review.

Parasites, including viruses, bacteria, fungi, protists, helminths, and arthropods, are ubiquitous in the animal kingdom. Consequently, hosts are frequently infected with more than one parasite species simultaneously. The assessment of such co-infections is of fundamental importance for disease ecology, but relevant studies involving non-domesticated animals have remained scarce. Many amphibians are in decline, and they generally have a highly diverse parasitic fauna. Here we review the literature reporting on field surveys, veterinary case studies, and laboratory experiments on co-infections in amphibians, and we summarize what is known about within-host interactions among parasites, which environmental and intrinsic factors influence the outcomes of these interactions, and what effects co-infections have on hosts. The available literature is piecemeal, and patterns are highly diverse, so that identifying general trends that would fit most host-multiparasite systems in amphibians is difficult. Several examples of additive, antagonistic, neutral, and synergistic effects among different parasites are known, but whether members of some higher taxa usually outcompete and override the effects of others remains unclear. The arrival order of different parasites and the time lag between exposures appear in many cases to fundamentally shape competition and disease progression. The first parasite to arrive can gain a marked reproductive advantage or induce cross-reaction immunity, but by disrupting the skin and associated defences (i.e., skin secretions, skin microbiome) and by immunosuppression, it can also pave the way for subsequent infections. Although there are exceptions, detrimental effects to the host are generally aggravated with increasing numbers of co-infecting parasite species. Finally, because amphibians are ectothermic animals, temperature appears to be the most critical environmental factor that affects co-infections, partly via its influence on amphibian immune function, partly due to its direct effect on the survival and growth of parasites. Besides their importance for our understanding of ecological patterns and processes, detailed knowledge about co-infections is also crucial for the design and implementation of effective wildlife disease management, so that studies concentrating on the identified gaps in our understanding represent rewarding research avenues.

Toxoplasma gondii in mollusks and cold-blooded animals: a systematic review.

Toxoplasma gondii (T. gondii) is known for its ability to infect warm-blooded vertebrates. Although T. gondii does not appear to parasitize cold-blooded animals, the occurrence of T. gondii infection in marine mammals raises concerns that cold-blooded animals (frogs, toad, turtles, crocodiles, snakes, and fish) and shellfish are potential sources of T. gondii. Therefore, this systematic review aimed to determine the prevalence of T. gondii in mollusks and cold-blooded animals worldwide. We searched PubMed, ScienceDirect, ProQuest, Scopus, and Web of Science from inception to 1 August 2020 for eligible papers in the English language and identified 26 articles that reported the prevalence of T. gondii in mollusks and cold-blooded animals. These articles were subsequently reviewed and data extracted using a standard form. In total, 26 studies [involving 9 cross-sectional studies including 2988 samples of cold-blooded animals (129 positive cases for T. gondii) and 18 cross-sectional studies entailing 13 447 samples of shellfish (692 positive cases for T. gondii)] were included in this study. Although this study showed that shellfish and cold-blooded animals could be potential sources of T. gondii for humans and other hosts that feed on them, further investigations are recommended to determine the prevalence of T. gondii in shellfish and cold-blooded animals.

Cryptic speciation among Tylodelphys spp.: the major helminth pathogens of fish and amphibians.

Larvae of Tylodelphys Diesing, 1950 are major digenean pathogens of fish and amphibians. Tylodelphys spp. may induce mass mortality of fish and increase their susceptibility to predation. Even though Tylodelphys spp. cause substantial damage to aquaculture systems, surprisingly little is known regarding the taxonomy of this commercially important genus with a limited number of visible autapomorphic identification features. The authors obtained the DNA sequences and analyzed the molecular phylogenetics of Tylodelphys spp. adults isolated from bird hosts of Czech origin and provide comparative measurements of the analyzed species. They identified a previously unknown species complex that is subject to cryptic speciation and was previously morphologically identified as Tylodelphys excavata (Rudolphi, 1803) sensu lato. This species complex consists of three morphologically similar but genetically well-separated species. Tylodelphys excavata sensu stricto remains the dominant Tylodelphys isolated from Ciconia ciconia, which also serves as a satellite host of Tylodelphys circibuteonis Odening, 1962, which is the resurrected species for which birds of prey serve as core hosts. The authors describe Tylodelphys nigriciconis sp. n. Heneberg & Sitko as a new species identified in Ciconia nigra. By providing the first sequences of Tylodelphys podicipina Kozicka and Niewiadomska, 1960, they also show that Tylodelphys immer Dubois, 1961 is a junior synonym of T. podicipina. Further research is needed to match the provided molecular data with the DNA of larval Tylodelphys from outbreaks in commercially exploited fish species.

Biodiversity of the Coccidia (Apicomplexa: Conoidasida) in vertebrates: what we know, what we do not know, and what needs to be done.

Over the last two decades my colleagues and I have assembled the literature on a good percentage of most of the coccidians (Conoidasida) known, to date, to parasitise: Amphibia, four major lineages of Reptilia (Amphisbaenia, Chelonia, Crocodylia, Serpentes), and seven major orders in the Mammalia (Carnivora, Chiroptera, Lagomorpha, Insectivora, Marsupialia, Primates, Scandentia). These vertebrates, combined, comprise about 15,225 species; only about 899 (5.8%) of them have been surveyed for coccidia and 1,946 apicomplexan valid species names or other forms are recorded in the literature. Based on these compilations and other factors, I extrapolated that there yet may be an additional 31,381 new apicomplexans still to be discovered in just these 12 vertebrate groups. Extending the concept to all of the other extant vertebrates on Earth; i.e. lizards (6,300 spp.), rodents plus 12 minor orders of mammals (3,180 spp.), birds (10,000 spp.), and fishes (33,000 spp.) and, conservatively assuming only two unique apicomplexan species per each vertebrate host species, I extrapolate and extend my prediction that we may eventually find 135,000 new apicomplexans that still need discovery and to be described in and from those vertebrates that have not yet been examined for them! Even doubling that number is a significant underestimation in my opinion.

The cost of travel: How dispersal ability limits local adaptation in host-parasite interactions.

Classical theory suggests that parasites will exhibit higher fitness in sympatric relative to allopatric host populations (local adaptation). However, evidence for local adaptation in natural host-parasite systems is often equivocal, emphasizing the need for infection experiments conducted over realistic geographic scales and comparisons among species with varied life history traits. Here, we used infection experiments to test how two trematode (flatworm) species (Paralechriorchis syntomentera and Ribeiroia ondatrae) with differing dispersal abilities varied in the strength of local adaptation to their amphibian hosts. Both parasites have complex life cycles involving sequential transmission among aquatic snails, larval amphibians and vertebrate definitive hosts that control dispersal across the landscape. By experimentally pairing 26 host-by-parasite population infection combinations from across the western USA with analyses of host and parasite spatial genetic structure, we found that increasing geographic distance-and corresponding increases in host population genetic distance-reduced infection success for P. syntomentera, which is dispersed by snake definitive hosts. For the avian-dispersed R. ondatrae, in contrast, the geographic distance between the parasite and host populations had no influence on infection success. Differences in local adaptation corresponded to parasite genetic structure; although populations of P. syntomentera exhibited ~10% mtDNA sequence divergence, those of R. ondatrae were nearly identical (<0.5%), even across a 900 km range. Taken together, these results offer empirical evidence that high levels of dispersal can limit opportunities for parasites to adapt to local host populations.

Distribution modeling of Amblyomma rotundatum and Amblyomma dissimile in Brazil: estimates of environmental suitability.

The number of reports of tick parasitism in amphibians and reptiles has increased over the past few years, including new host and location records for Amblyomma rotundatum and Amblyomma dissimile. However, knowledge of the geographical distribution remains incomplete, and in many regions of Brazil, the presence of these vectors has not been investigated. Several candidate models were built using a correlative maximum entropy approach, and best-fitting models were selected based on statistical significance, predictive ability, and complexity based on current climatic trends and future projected climate changes. Final models showed a good ability to discriminate A. rotundatum and A. dissimile current and future potential distributions. The entire country had higher predicted suitability for A. rotundatum while A. dissimile was mainly restricted to the Amazon and Pantanal biomes. A. rotundatum is a species with enormous potential for dissemination in the next decades, potentially through the legal and illegal transport of reptiles and amphibians. The proposed model is useful for targeting surveillance efforts increasing the efficiency and accuracy of future ecological research and tick management efforts.

Synopsis of the species of the genus Zschokkella Auerbach, 1910 (Myxozoa: Bivalvulida: Myxidiidae).

A synopsis of the species of Zschokkella Auerbach, 1910 (Myxozoa: Bivalvulida: Myxidiidae) is presented, including 94 nominal species from piscine hosts and 3 additional nominal species from amphibian or reptilian hosts. The most relevant morphological and morphometric features of both myxospores and trophozoite stages (when available) are presented for each species in tabulated format.

Dracunculus Species in Meso-mammals from Georgia, United States, and Implications for the Guinea Worm Eradication Program in Chad, Africa.

The prevalence and diversity of parasitic nematodes in wildlife have been well studied for certain species, yet for others considerable gaps in knowledge exist. The parasitic nematode Dracunculus insignis infects North American wildlife, and past research on this species has led to an increased understanding of the potential host diversity and transmission of the closely related human Guinea worm, Dracunculus medinensis (which is currently the focus of a global eradication program). Many definitive hosts have been documented for D. insignis; however, the life cycle has been studied only in laboratories, and only a single phylogenetic study has been conducted on D. insignis (from Canada). The goals of the present study were to investigate the prevalence of infections with Dracunculus species among wildlife at a single site (Di-Lane plantation) in the southeastern United States, evaluate the genetic diversity of parasites at this site, and investigate potential paratenic hosts that may be involved in transmission. Over 3 yr, we sampled 228 meso-mammals, reporting an overall prevalence of infection with Dracunculus insignis of 20% (46/228). Amphibians and fish were sampled in the same geographic area as infected meso-mammals. Dracunculus insignis third-stage larvae were recovered from 2 different species of amphibians, but all fish sampled were negative. Phylogenetic analysis of the partial cytochrome c oxidase I (COI) gene showed very little diversity of Dracunculus at Di-Lane; however, we did recover a single nematode from a Virginia opossum (Didelphis virginiana) that falls outside of the D. insignis clade, more closely aligns with Dracunculus lutrae, and may represent an undescribed species. This work documents the occurrence of D. insignis in frogs, a potential transmission pathway for D. insignis at a single geographic site in nature. When applied to the global Guinea Worm Eradication Program, and Chad, Africa, in particular, this work increases our knowledge of the potential role of aquatic animals in the transmission of Dracunculus species and informs on potential intervention strategies that may be applied to the eradication of Guinea worm in Africa.

Multilocus Metabarcoding of Terrestrial Leech Bloodmeal iDNA Increases Species Richness Uncovered in Surveys of Vertebrate Host Biodiversity.

Leech-derived invertebrate DNA (iDNA) has been successfully leveraged to conduct surveys of vertebrate host biodiversity across the Indo Pacific. However, this technique has been limited methodologically, typically only targeting mammalian 16S rDNA, or both 16S and vertebrate 12S rDNA for leech host determination. To improve the taxonomic richness of vertebrate host species in iDNA surveys, we re-analyze datasets from Bangladesh, Cambodia, China, and Madagascar through metabarcoding via next generation sequencing (NGS) of 12S, 16S (2 types, one designed to target mammals and the other, residual eDNA), nicotinamide adenine dinucleotide hydride dehydrogenase 2 (ND2), and cytochrome c oxidase subunit 1 (COI). With our 5 primer sets, we identify 41 unique vertebrate hosts to the species level, among 1,200 leeches analyzed, along with an additional 13 taxa to the family rank. Within our 41 taxa, we note that adding ND2 and COI loci increased species richness detection by 25%. NGS has emerged as more efficient than Sanger sequencing for large scale metabarcoding applications and, with the decline in cost of NGS, our pooled sample multilocus protocol is an attractive option for iDNA biodiversity surveys.

Amphibian Host and Skin Microbiota Response to a Common Agricultural Antimicrobial and Internal Parasite.

Holistic approaches that simultaneously characterize responses of both microbial symbionts and their hosts to environmental shifts are imperative to understanding the role of microbiotas on host health. Using the northern leopard frog (Lithobates pipiens) as our model, we investigated the effects of a common trematode (family Echinostomatidae), a common agricultural antimicrobial (Sulfadimethoxine; SDM), and their interaction on amphibian skin microbiota and amphibian health (growth metrics and susceptibility to parasites). In the trematode-exposed individuals, we noted an increase in alpha diversity and a shift in microbial communities. In the SDM-treated individuals, we found a change in the composition of the skin microbiota similar to those induced by the trematode treatment. Groups treated with SDM, echinostomes, or a combination of SDM and echinostomes, had higher relative abundances of OTUs assigned to Flavobacterium and Acinetobacter. Both of these genera have been associated with infectious disease in amphibians and the production of anti-pathogen metabolites. Similar changes in microbial community composition between SDM and trematode exposed individuals may have resulted from stress-related disruption of host immunity. Despite changes in the microbiota, we found no effect of echinostomes and SDM on host health. Given the current disease- and pollution-related threats facing amphibians, our study highlights the need to continue to evaluate the influence of natural and anthropogenic stressors on host-associated microbial communities.

Selected Wildlife Trematodes.

The trematodes are a species-rich group of parasites, with some estimates suggesting that there are more than 24,000 species. However, recent interests on the biology of trematodes of wildlife indicate that the taxonomic status and nomenclature of many of the previous species descriptions of wildlife trematodes throughout the world are confusing and difficult to decipher. In this chapter, we review work on selected trematodes of amphibians, birds, mammals, and their snail intermediate hosts, in the hope of providing a tool kit on how to study trematodes of wildlife. We provide a brief introduction to each group of wildlife trematodes, followed by some examples of the challenges each group of trematodes has relative to the goal of their identification and understanding of their biology and interactions with their wildlife hosts.

Biological Concepts for the Control of Aquatic Zoosporic Diseases.

Aquatic zoosporic diseases are threatening global biodiversity and ecosystem services, as well as economic activities. Current means of controlling zoosporic diseases are restricted primarily to chemical treatments, which are usually harmful or likely to be ineffective in the long term. Furthermore, some of these chemicals have been banned due to adverse effects. As a result, there is a need for alternative methods with minimal side-effects on the ecosystem or environment. Here, we integrate existing knowledge of three poorly interconnected areas of disease research - amphibian conservation, aquaculture, and plankton ecology - and arrange it into seven biological concepts to control zoosporic diseases. These strategies may be less harmful and more sustainable than chemical approaches. However, more research is needed before safe application is possible.

Patterns of Clinostomum marginatum infection in fishes and amphibians: integration of field, genetic, and experimental approaches.

Digenetic trematodes of the genus Clinostomum are cosmopolitan parasites infecting fishes, amphibians, reptiles, and snails as intermediate hosts. Despite the broad geographical distribution of this genus, debate about the number of species and how they vary in host use has persisted. To better understand patterns of infection among host species and across life stages, we used large-scale field surveys and molecular tools to examine five species of amphibians and seven species of fishes from 125 California ponds. Among the 12,360 examined hosts, infection was rare, with an overall prevalence of 1.7% in amphibians and 9.2% in fishes. Molecular evidence indicated that both groups were infected with Clinostomum marginatum. Using generalized linear mixed effects models, host species identity and host life stage had a strong influence on infection status, such that Lepomis cyanellus (green sunfish) (49.3%) and Taricha granulosa (rough skinned newt) (9.2%) supported the highest overall prevalence values, whereas adult amphibians tended to have a higher prevalence of infection relative to juveniles (13.3% and 2.5%, respectively). Experimentally, we tested the susceptibility of two amphibian hosts (Pseudacris regilla [Pacific chorus frog] and Anaxyrus boreas [western toad]) to varying levels of cercariae exposure and measured metacercariae growth over time. Pseudacris regilla was 1.3× more susceptible to infection, while infection success increased with cercariae exposure dose for both species. On average, metacarcariae size increased by 650% over 20 days. Our study highlights the importance of integrating field surveys, genetic tools, and experimental approaches to better understand the ecology of host-parasite interactions.

Direct and indirect effects of a common cyanobacterial toxin on amphibian-trematode dynamics.

Wildlife diseases are emerging at unprecedented rates. While there are likely several factors at play, human-mediated environmental alterations may play a significant role. Of growing interest is the effect that microcystin-LR (MC-LR), a cyanotoxin, may have on disease outcomes. In this study, using an amphibian-trematode model we examined (1) the lethal effects of MC-LR on cercariae of trematodes; (2) the sublethal effects of MC-LR exposure on the ability for trematodes to infect green frog tadpoles; and (3) the sublethal effects of MC-LR on green frog tadpole susceptibility to trematodes. We found that environmentally-relevant concentrations of MC-LR at 50, 100, and 500 µg L-1 increased cercariae rate of mortality (LC50-14h = 134.24 µg L-1). However, sublethal exposure of trematodes to 2 and 10 µg L-1 MC-LR did not alter their infectivity. Conversely, sublethal exposure of tadpoles to 2 µg L-1 increased their susceptibility to trematodes by 147%. However, 10 µg L-1 of MC-LR did not affect tadpole susceptibility to trematodes, indicating a non-linear response to sublethal MC-LR exposure. Overall, our findings suggest that high concentrations of MC-LR (≥50 µg L-1) have the potential to limit trematode transmission to amphibian hosts through MC-LR-induced mortality. However, at lower concentrations (<10 µg L-1) MC-LR's effect on tadpole-cercariae disease outcome is likely driven by its effect on the tadpole host. Collectively, this work highlights the need to consider how toxicants influence both host and parasite at multiple concentrations to better understand the impacts of cyanotoxins on disease dynamics.

Endocrine and immune responses of larval amphibians to trematode exposure.

In nature, multiple waves of exposure to the same parasite are likely, making it important to understand how initial exposure or infection affects subsequent host infections, including the underlying physiological pathways involved. We tested whether experimental exposure to trematodes (Echinostoma trivolvis or Ribeiroia ondatrae) affected the stress hormone corticosterone (known to influence immunocompetence) in larvae representing five anuran species. We also examined the leukocyte profiles of seven host species after single exposure to R. ondatrae (including four species at multiple time points) and determined if parasite success differed between individuals given one or two challenges. We found strong interspecific variation among anuran species in their corticosterone levels and leukocyte profiles, and fewer R. ondatrae established in tadpoles previously challenged, consistent with defense "priming." However, exposure to either trematode had only weak effects on our measured responses. Tadpoles exposed to E. trivolvis had decreased corticosterone levels relative to controls, whereas those exposed to R. ondatrae exhibited no change. Similarly, R. ondatrae exposure did not lead to appreciable changes in host leukocyte profiles, even after multiple challenges. Prior exposure thus influenced host susceptibility to trematodes, but was not obviously associated with shifts in leukocyte counts or corticosterone, in contrast to work with microparasites.

Amblyomma ticks infesting amphibians and Squamata reptiles from the lower Amazon region, Brazil.

Amphibians and Squamata reptiles belonging to a zoological collection were screened for ectoparasites, which were removed from the hosts and identified using morphological keys. Descriptive statistics and analysis of the association between the parasite and host characteristics (taxonomic group, capture location and habitat) were done. Among the 1256 animals examined (319 amphibians and 937 reptiles), 86 individuals were parasitized, corresponding to a frequency of 6.9% (6.6% reptiles and 7.5% amphibians). Ticks in the adult and nymph stages were identified to the species level; all of them belonged to the species Amblyomma dissimile. The larvae were identified to the genus level and were all Amblyomma sp. In total 69 larvae, 28 nymphs and eight adults were found. The most parasitized species was the frog Rhinella major: 24 parasitized animals of 65 examined (36.9%). There was a difference (P < 0.001) between parasitism by ticks of the genus Amblyomma with regard to the habitat of capture of the parasitized animal, with a higher parasitism rate in hosts that inhabited open areas as compared to animals ensconced in forest areas and edges of forests. New tick-host associations are given.