Centrorhynchus spp. (Acanthocephala) in South America: new anuran record and checklist of vertebrate hosts.

The aim of this study was to record Centrorhynchus sp. associated with the exotic species Aquarana catesbeiana (bullfrog) in southern Brazil and to present a checklist of vertebrate hosts in South America. Twenty-nine adults and juveniles of A. catesbeiana were collected in Capão do Leão, state of Rio Grande do Sul, Brazil, between October 2019 and December 2020. We found 275 specimens of Centrorhynchus sp. cystacanths in the stomach musculature and coelomic cavity of 55.1% of hosts (16). There was no significant differences in the prevalence and mean intensity of infection with cystacanths when compared males and females of A. catesbeiana. The prevalence was significantly higher in adults than in juveniles. The checklist presents 106 species of vertebrate hosts and 14 taxa of Centrorhynchus recorded in nine South American countries. Avian were the main definitive hosts of Centrorhynchus spp. and snakes Dipsadidae, anurans Hylidae and Leptodactylidae the main paratenic hosts in South America. This is the first record of Centrorhynchus cystacanths in A. catesbeiana in the South America. The study provides tools to help understand the parasitic relationships between species of Centrorhynchus and A. catesbeiana and other hosts in areas where bullfrog have been introduced.

Pattern of anuran infection by acanthocephalans from the Cerrado, Northeastern Brazil with a summary for South America.

In Brazil, acanthocephalans parasitise anurans in several biomes. In the present study, we performed an analysis of acanthocephalan infections across 175 anuran individuals from the Cerrado biome, belonging to ten species: Boana raniceps, Pithecopus hypochondrialis, Scinax fuscomarginatus, Scinax x-signatus, Leptodactylus pustulatus, Leptodactylus macrosternum, Leptodactylus vastus, Physalaemus cuvieri, Adenomera hylaedactyla, and Elachistocleis piauiensis. We also verified the specificity of the parasites using the STD* index. Additionally, we conducted a survey of acanthocephalan infection in anurans in South America. The studied assemblage in the Brazilian Cerrado presented 57 parasitised hosts of 175 specimens (overall prevalence: 32.6%). In total, 437 acanthocephalans cystacanths were recorded, among which 286 presented the same morphotype but could not be identified, 148 belonged to the genus Centrorhynchus, and three belonged to Oncicola. Unidentified acanthocephalans had a higher prevalence in L. vastus (53.85%) and the highest intensity was in L. pustulatus (17±16). The highest prevalence of Centrorhynchus sp. was in the species S. fuscomarginatus (28.57%), while the highest intensity was observed in L. vastus (111). The taxon Oncicola sp. it had a prevalence of 3.23% and an intensity of 3 only in S. x-signatus. The highest specificity was recorded for Oncicola sp. (STD*= 1), whereas the lowest was found in Centrorhynchus sp. (STD*= 2.21). Finally, according to the survey for South America, we found ten records of acanthocephalan taxa parasitizing 58 species of anurans distributed in seven countries (Brazil with the most records).

Putting infection on the map: Using heatmaps to characterise within- and between-host distributions of trematode metacercariae.

The location of parasites within individual hosts is often treated as a static trait, yet many parasite species can occur in multiple locations or organs within their hosts. Here, we apply distributional heat maps to study the within- and between-host infection patterns for four trematodes (Alaria marcianae, Cephalogonimus americanus, Echinostoma spp. and Ribeiroia ondatrae) within the amphibian hosts Pseudacris regilla and two species of Taricha. We developed heatmaps from 71 individual hosts from six locations in California, which illustrate stark differences among parasites both in their primary locations within amphibian hosts as well as their degree of location specificity. While metacercariae (i.e., cysts) of two parasites (C. americanus and A. marcianae) were relative generalists in habitat selection and often occurred throughout the host, two others (R. ondatrae and Echinostoma spp.) were highly localised to a specific organ or organ system. Comparing parasite distributions among these parasite taxa highlighted locations of overlap showing potential areas of interactions, such as the mandibular inner dermis region, chest and throat inner dermis and the tail reabsorption outer epidermis. Additionally, the within-host distribution of R. ondatrae differed between host species, with metacercariae aggregating in the anterior dermis areas of newts, compared with the posterior dermis area in frogs. The ability to measure fine-scale changes or alterations in parasite distributions has the potential to provide further insight about ecological questions concerning habitat preference, resource selection, host pathology and disease control.

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.

Description of a new species of Aplectana (Nematoda: Ascaridomorpha: Cosmocercidae) using an integrative approach and preliminary phylogenetic study of Cosmocercidae and related taxa.

BACKGROUND: Nematodes of the family Cosmocercidae (Ascaridomorpha: Cosmocercoidea) are mainly parasitic in the digestive tract of various amphibians and reptiles worldwide. However, our knowledge of the molecular phylogeny of the Cosmocercidae is still far from comprehensive. The phylogenetic relationships between Cosmocercidae and the other two families, Atractidae and Kathlaniidae, in the superfamily Cosmocercoidea are still under debate. Moreover, the systematic position of some genera within Cosmocercidae remains unclear. METHODS: Nematodes collected from Polypedates megacephalus (Hallowell) (Anura: Rhacophoridae) were identified using morphological (light and scanning electron microscopy) and molecular methods [sequencing the small ribosomal DNA (18S), internal transcribed spacer 1 (ITS-1), large ribosomal DNA (28S) and mitochondrial cytochrome c oxidase subunit 1 (cox1) target regions]. Phylogenetic analyses of cosmocercoid nematodes using 18S + 28S sequence data were performed to clarify the phylogenetic relationships of the Cosmocercidae, Atractidae and Kathlaniidae in the Cosmocercoidea and the systematic position of the genus Aplectana in Cosmocercidae. RESULTS: Morphological and genetic evidence supported the hypothesis that the nematode specimens collected from P. megacephalus represent a new species of Aplectana (Cosmocercoidea: Cosmocercidae). Our phylogenetic results revealed that the Cosmocercidae is a monophyletic group, but not the basal group in Cosmocercoidea as in the traditional classification. The Kathlaniidae is a paraphyletic group because the subfamily Cruziinae within Kathlaniidae (including only the genus Cruzia) formed a seperate lineage. Phylogenetic analyses also showed that the genus Aplectana has a closer relationship to the genus Cosmocerca in Cosmocercidae. CONCLUSIONS: Our phylogenetic results suggested that the subfamily Cruziinae should be moved from the hitherto-defined family Kathlaniidae and elevated as a separate family, and the genus Cosmocerca is closely related to the genus Aplectana in the family Cosmocercidae. The present study provided a basic molecular phylogenetic framework for the superfamily Cosmocercoidea based on 18S + 28S sequence data for the first time to our knowledge. Moreover, a new species, A. xishuangbannaensis n. sp., was described using integrative approach.

Zygocaulus nagoensis n. gen. and n. sp. (Trichostrongyloidea: Dictyocaulidae), a Peculiar Bursate Nematode Collected from an Alien Frog, Polypedates leucomystax, in Nago, Okinawa Island, Japan.

A peculiar bursate nematode, Zygocaulus nagoensis n. gen. and n. sp. (Trichostrongyloidea: Dictyocaulidae: Mertensinematinae), was described from an alien frog, Polypedates leucomystax (Anura: Rhacophoridae), collected on Okinawa Island, Japan. It is related to Mertensinema and Borrellostrongylus, the only hitherto known genera of Mertensinematinae, but is readily distinguished from them by having only 2 pairs of lateral rays and simple distal ends of the dorsal ray branches.

Balantidium grimi n. sp. (Ciliophora, Litostomatea), a new species inhabiting the rectum of the frog Quasipaa spinosa from Lishui, China.

Balantidium grimi n. sp. is described from the rectum of the frog Quasipaa spinosa (Amphibia, Dicroglossidae) from Lishui, Zhejiang Province, China. The new species is described by both light microscopy (LM) and scanning electron microscopy (SEM), and a molecular phylogenetic analysis is also presented. This species has unique morphological features in that the body shape is somewhat flattened and the vestibulum is "V"-shaped, occupying nearly 3/8 to 4/7 of the body length. Only one contractile vacuole, situated at the posterior body, was observed. The phylogenetic analysis based on SSU-rDNA indicates that B. grimi groups together with B. duodeni and B. entozoon. In addition, the genus Balantidium is clearly polyphyletic.

Helminths of Dermatonotus muelleri (Anura: Microhylidae) from Northeastern Brazil.

The helminth fauna associated with Muller's termite frog, Dermatonotus muelleri, from the southern region of Ceará State, Brazil, was studied. The species richness was 6 helminth taxa, including cystacanths of Acanthocephala and 5 nematode species: Aplectana membranosa, Parapharyngodon silvoi, Raillietnema spectans, larvae of Physaloptera sp., and an unidentified nematode. The overall prevalence was 88.6%, with an average intensity of infection of 123.7 ± 26.3. The nematode Raillietnema spectans presented the highest prevalence and was the most abundant ( d = 0.670). Host body size did not influence the intensity of infection nor the richness of helminth species. This study increases the body of knowledge about the diversity of helminth fauna associated with Dermatonotus muelleri from northeastern Brazil, extending the record of hosts and the geographic distribution of these helminth species.

High prevalence of chigger mite infection in a forest-specialist frog with evidence of parasite-related granulomatous myositis.

Amphibians are hosts for a wide variety of micro- and macro-parasites. Chigger mites from the Hannemania genus are known to infect a wide variety of amphibian species across the Americas. In Chile, three species (H. pattoni, H. gonzaleacunae and H. ortizi) have been described infecting native anurans; however, neither impacts nor the microscopic lesions associated with these parasites have been described. Here, we document 70% prevalence of chigger mite infection in Eupsophus roseus and absence of infection in Rhinoderma darwinii in the Nahuelbuta Range, Chile. Additionally, we describe the macroscopic and microscopic lesions produced by H. ortizi in one of these species, documenting previously undescribed lesions (granulomatous myositis) within the host's musculature. These findings highlight that further research to better understand the impacts of chigger mite infection on amphibians is urgently required in Chile and elsewhere.

Choledocystus elegans (Digenea: Plagiorchiidae) of Leptodactylus paraensis (Amphibia: Leptodactylidae) from the Brazilian Amazon / Choledocystus elegans (Digenea: Plagiorchiidae) de Leptodactylus paraensis (Amphibia: Leptodactylidae) da Amazônia brasileira

Abstract The trematodes are parasites of the several vertebrates including amphibians, however the knowledge about of the taxonomy these parasites is still confuse. The trematode Choledocystus elegans was found in the small intestine of the Leptodactylus paraensis in eastern Amazon and presents the following characteristics: several pointed tegumentary spines, papillae on the outer and inner edges of the oral and ventral suckers, a round, well-developed cirrus sac, a well-developed cirrus, oblique testicles, a ovary right side, uterine loops extending between the testicles, follicular vitellaria distributed throughout the body, starting at the genital pore region and caeca close the end of the body. For the first time, this study identified C. elegans parasitizing L. paraensis and describes morphological aspects never characterized using light and scanning electron microscopy.

Resumo Os trematodas são parasitas de vários grupos de vertebrados incluindo os anfíbios, contudo o que se conhece sobre a taxonomia destes parasitas ainda é confusa. O trematoda Choledocystus elegans foi encontrado no intestino delgado de Leptodactylus paraensis na Amazônia oriental e apresenta as seguintes características: vários espinhos pontiagudos no tegumento, papilas nas bordas externa e interna das ventosas orais e ventrais, bolsa do cirro bem desenvolvida contendo um cirro desenvolvido, testículos oblíquos, ovário destro, alças uterinas que se estendem entre os testículos, folículos vitelínicos distribuída por toda a lateral do corpo, começando ao nível do poro genital e cecos intestinais chegando até a região posterior do corpo do helminto. Pela primeira vez, este estudo identificou C. elegans parasitando L. paraensis e descreve aspectos morfológicos nunca caracterizados usando microscopia de luz comum e eletrônica de varredura.

Helminth fauna of Leptodactylus syphax (Anura: Leptodactylidae) from Caatinga biome, northeastern Brazil.

Leptodactylus syphax is distributed in central, southeastern and northeastern Brazil, eastern Bolivia and southern Paraguay, occupying open areas and rock outcrops, in rock cavities and termite burrows. We collected 21 frogs from the Caatinga region of the state of Ceará, northeastern Brazil, and 7,021 helminths were recovered from 18 of these hosts (overall prevalence = 85.7%). Six helminth taxa were recovered, as follows: Aplectana membranosa (n = 3,756); Schrankiana formosula (n = 3,176); larvae of Physaloptera sp. (n = 43); unidentified nematode larvae (n = 7); digenean metacercariae of Lophosicyadiplostomum sp. (n = 2); and cystacanths of Acanthocephala (n = 37). The similarity of helminth composition between L. syphax from the Caatinga and other species of the L. fuscus group showed that some anurans were clustered according to parasite species and others according to geographic locality. This study presents new helminth records for the Neotropical region, thus helping in understanding the pattern of species distribution, and it increases the knowledge of parasites associated with amphibians.

Helminth fauna of Leptodactylus syphax (Anura: Leptodactylidae) from Caatinga biome, northeastern Brazil / Helmintofauna de Leptodactylus syphax (Anura: Leptodactylidae) do bioma da Caatinga, Nordeste do Brasil

Abstract Leptodactylus syphax is distributed in central, southeastern and northeastern Brazil, eastern Bolivia and southern Paraguay, occupying open areas and rock outcrops, in rock cavities and termite burrows. We collected 21 frogs from the Caatinga region of the state of Ceará, northeastern Brazil, and 7,021 helminths were recovered from 18 of these hosts (overall prevalence = 85.7%). Six helminth taxa were recovered, as follows: Aplectana membranosa (n = 3,756); Schrankiana formosula (n = 3,176); larvae of Physaloptera sp. (n = 43); unidentified nematode larvae (n = 7); digenean metacercariae of Lophosicyadiplostomum sp. (n = 2); and cystacanths of Acanthocephala (n = 37). The similarity of helminth composition between L. syphax from the Caatinga and other species of the L. fuscus group showed that some anurans were clustered according to parasite species and others according to geographic locality. This study presents new helminth records for the Neotropical region, thus helping in understanding the pattern of species distribution, and it increases the knowledge of parasites associated with amphibians.

Resumo Leptodactylus syphax está distribuída na região central, Sudeste e Nordeste do Brasil, Leste da Bolívia e Sul do Paraguai, ocupando áreas abertas e afloramentos rochosos, cavidades rochosas ou de cupins. Foram coletadas 21 rãs oriundas da região de Caatinga do Ceará, nordeste brasileiro, e 7.021 helmintos foram recuperados em 18 hospedeiros (prevalência geral = 85,7%). Seis taxa de helmintos foram recuperados, como segue: Aplectana membranosa (n = 3.756), Schrankiana formosula (n = 3.176), larvas de Physaloptera sp. (n = 43), larvas de nematódeos não identificado (n = 7), metacercárias de Lophosicyadiplostomum sp. (n = 2), e cistacantos de Acantocéfalos (n = 37). A similaridade da composição de helmintos entre L. syphax da Caatinga e outras espécies do grupo L. fuscus mostrou que alguns anuros foram agrupados de acordo com a espécie do parasita e outros de acordo com a localidade geográfica. Este estudo apresenta novos registros de helmintos para a região Neotropical, ajudando na compreensão do padrão de distribuição das espécies e aumenta o conhecimento sobre os parasitas associados a anfíbios.

Role of Antimicrobial Peptides in Amphibian Defense Against Trematode Infection.

Antimicrobial peptides (AMPs) contribute to the immune defenses of many vertebrates, including amphibians. As larvae, amphibians are often exposed to the infectious stages of trematode parasites, many of which must penetrate the host's skin, potentially interacting with host AMPs. We tested the effects of the natural AMPs repertoires on both the survival of trematode infectious stages as well as their ability to infect larval amphibians. All five trematode species exhibited decreased survival of cercariae in response to higher concentrations of adult bullfrog AMPs, but no effect when exposed to AMPs from larval bullfrogs. Similarly, the use of norepinephrine to remove AMPs from larval bullfrogs, Pacific chorus frogs, and gray treefrogs had only weak (gray treefrogs) or non-significant (other tested species) effects on infection success by Ribeiroia ondatrae. We nonetheless observed strong differences in parasite infection as a function of both host stage (first- versus second-year bullfrogs) and host species (Pacific chorus frogs versus gray treefrogs) that were apparently unrelated to AMPs. Taken together, our results suggest that AMPs do not play a significant role in defending larval amphibians against trematode cercariae, but that they could be one mechanism helping to prevent infection of post-metamorphic amphibians, particularly for highly aquatic species.

Species complexes and phylogenetic lineages of Hoferellus (Myxozoa, Cnidaria) including revision of the genus: A problematic case for taxonomy.

BACKGROUND: Myxozoans are metazoan parasites whose traditional spore morphology-based taxonomy conflicts DNA based phylogenies. Freshwater species of the genus Hoferellus are parasites of the excretory system, with several members infecting food and ornamental fish species, as well as amphibians. This study aims to increase our understanding of their molecular diversity and development, aspects about which little is known, and to generate a molecular diagnostic tool to discriminate between different pathogenic and non-pathogenic Hoferellus spp. METHODS: SSU and ITS rDNA phylogeny, along with morphological descriptions using light and electron microscopy were used to identify and characterize Hoferellus species collected from the urinary system of fishes and frogs. A PCR-based diagnostic assay was designed to differentiate between cryptic Hoferellus spp in cyprinid fishes commonly cultured in Central Europe. RESULTS: Our phylogenetic results separate the species of Hoferellus into two phylogenetic sublineages which are indistinguishable on the basis of generic morphological traits: 1) The Hoferellus sensu stricto sublineage, which is composed of the type species Hoferellus cyprini, Hoferellus carassii and a cryptic species, Hoferellus sp. detected only molecularly in common carp. 2) The Hoferellus sensu lato sublineage into which the new species we described in this study, Hoferellus gnathonemi sp. n. from the kidney of the elephantnose fish and Hoferellus anurae from reed frogs, are placed together with Hoferellus gilsoni previously sequenced from European eel. Apart from phylogenetic analyses, we also provide novel ultrastructural data on the phagocytotic nature of some Hoferellus plasmodia and on the elusive intracellular stages ascribed to the presporogonic development of this genus. CONCLUSIONS: We provide molecular evidence of the polyphyly of the genus Hoferellus and provide novel morphological details of its members. Based on the presented data, we revise and propose emendation of the genus Hoferellus.

Dero (Allodero) lutzi Michaelsen, 1926 (Oligochaeta: Naididae) associated with Scinax fuscovarius (Lutz, 1925) (Anura: Hylidae) from Semi-deciduous Atlantic Rain Forest, southern Brazil / Dero (Allodero) lutzi Michaelsen, 1926 (Oligochaeta: Naididae) associado com Scinax fuscovarius (Lutz, 1925) (Anura: Hylidae) na Floresta Estacional Semidecidual, Sul do Brasil

Amphibians are hosts for a wide variety of ecto- and endoparasites, such as protozoans and parasitic worms. Naididae is a family of Oligochaeta whose species live on a wide range of substrates, including mollusks, aquatic macrophytes, sponges, mosses, liverworts, and filamentous algae. However, some species are known as endoparasitic from vertebrates, such as Dero (Allodero) lutzi, which is parasitic of the urinary tracts of frogs, but also have a free-living stage. Specimens in the parasitic stage lack dorsal setae, branchial fossa, and gills. Here we report the occurrence of D. (A.) lutzi associated with anuran Scinax fuscovarius from Semi-deciduous Atlantic Rain Forest in southern Brazil. The study took place at the Caiuá Ecological Station, Diamante do Norte, Paraná, southern Brazil. Seven specimens of S. fuscovarius were examined for parasites but only one was infected. Parasites occurred in ureters and urinary bladder. Previous records of this D. (A.) lutzi include the Brazilian States of Santa Catarina, São Paulo, Rio de Janeiro, and Minas Gerais, as well as Cuba and North America. This is a new locality record for this species in Brazil. Reports of Dero (Allodero) lutzi are rare, due to difficulty of observation, and such events are restricted only the fortuitous cases. It is important to emphasize the necessity of future studies, which are fundamental to the understanding of biological and ecological aspects of this species.

Anfíbios são hospedeiros para uma grande variedade de ecto- e endoparasitos, tais como os protozoários e vermes parasitos. Naididae é uma família de Oligochaeta cujas espécies vivem em uma ampla variedade de substratos, incluindo moluscos, macrófitas aquáticas, esponjas, musgos, e algas hepáticas e filamentosas. No entanto, algumas espécies são conhecidas como endoparasitos de vertebrados, como Dero (Allodero) lutzi, o qual é um parasito do trato urinário de anfíbios, mas que também possui uma fase de vida livre. Espécimes no estágio parasitário não possuem cerdas dorsais, fossa branquial, e brânquias. Relatamos a ocorrência de D. (A.) lutzi associado ao anuro Scinax fuscovarius na Floresta Estacional Semidecidual no Sul do Brasil. O estudo foi realizado na Estação Ecológica do Caiuá, Diamante do Norte, Paraná, Sul do Brasil. Sete espécimes de S. fuscovarius foram examinados para parasitos, mas apenas um estava infectado. Os parasitos ocorreram nos ureteres e bexiga urinária. Registros anteriores de D. (A.) lutzi incluem os estados brasileiros de Santa Catarina, São Paulo, Rio de Janeiro e Minas Gerais, bem como Cuba e América do Norte. Este é o registro de uma nova localidade para esta espécie no Brasil. Registros de Dero (Allodero) lutzi são raros, devido à dificuldade de observação, e tais eventos são restritos apenas a casos fortuitos. É importante ressaltar a necessidade de futuros estudos, que são fundamentais para a compreensão de aspectos biológicos e ecológicos desta espécie.

Helminth parasite communities of allopatric populations of the frog Leptodactylus podicipinus from Pantanal, Brazil.

Several factors may influence the structure of parasite communities in amphibian hosts. In this study, we describe the helminth parasites of three allopatric populations of the frog Leptodactylus podicipinus and test whether host size and sex were determinants of the structure and composition of the helminth communities. One hundred and twenty-three anurans were collected from three different study sites within the Pantanal wetlands and surveyed for helminth parasites. We found 14 helminth taxa: 7 species of nematodes, 4 species of trematodes, 1 species of cestodes, 1 species of acanthocephalan and one unidentified cyst. Host sex and size did not cause significant differences in helminth abundance or richness. The structure of helminth communities from the three study sites varied in terms of species composition, abundance and diversity. Six out of 14 helminth taxa were found in the three localities. Among those, the nematodes Cosmocerca podicipinus and Rhabdias sp., the trematode Catadiscus propinquus and the helminth cyst showed significant differences in mean abundances. We suggest that such differences found among the three component communities are driven by biotic and abiotic factors operating locally. Moreover, these differences stress the importance of local conditions, such as hydrologic characteristics and landscape composition, on helminth community structure.

Protozoan parasites of four species of wild anurans from a local zoo in Malaysia.

The parasitic protozoan fauna in sixty-six anurans comprising of Duttaphrynus melanostictus, Phrynoidis juxtaspera, Hylarana erythraea and Polypedates leucomystax collected from Zoo Negara Malaysia was investigated. The distribution and prevalence rate of parasitic species in the digestive tract and blood were examined. Seven species of intestinal protozoa (Opalina ranarum, Cepedea dimidiata, Nycthetorus cordiformis, Entamoeba ranarum, Iodamoeba butschlii, Endamoeba blattae, and Tritrichomonas sp.) and two species of blood protozoa (Lankesterella sp. and Trypanosoma sp.) were recorded. Opalina ranarum was the most common protozoan found in the rectum and intestine (prevalence rate: 34.8%) infecting all host species, with P. juxtaspera heavily infected with the parasite, whereas Tritrichomonas sp. was the least prevalent intestinal species infecting only D. melanostictus. Both Lankesterella sp. and Trypanosoma sp. were found in the blood of H. erythraea.

Comparative pathology and ecological implications of two myxosporean parasites in native Australian frogs and the invasive cane toad.

Myxosporean parasites Cystodiscus axonis and C. australis are pathogens of native and exotic Australian frog species. The pathology and ecological outcomes of infection with these parasites were investigated in this study. Gliosis was correlated to Cystodiscus axonis plasmodia in the brains of (9/60) tadpoles and (3/9) adult endangered Green and golden bell frogs using ordinal regression. Severe host reactions to C. axonis (haemorrhage, necrosis, and vasulitis) were observed in the brains of threatened Southern bell frogs (8/8), critically endangered Booroolong frogs (15/44) and Yellow spotted bell frogs (3/3). Severe brain lesions were associated with behavioural changes, neurological dysfunction, and spontaneous death. Both C. axonis and C. australis develop in the bile ducts of tadpoles, the plasmodia were significantly associated with biliary hyperplasia, inflammation and the loss of hepatocytes in (34/72) Green and golden bell frog tadpoles using ordinal regression. These lesions were so severe that in some cases 70% of the total liver was diseased. Normal liver function in tadpoles is necessary for metamorphosis, metabolism, and immune function. We postulate that this extensive liver damage would have significant host health impacts. Severe hepatic myxosporidiosis was more prevalent in tadpoles examined in autumn and winter (overwintered), suggestive of delayed metamorphosis in infected tadpoles, which would have serious flow-on effects in small populations. We compared the sensitivity of histopathology and species-specific PCR in the detection of C. australis and C. axonis. PCR was determined to be the most sensitive method (detection limit 1 myxospore equivalent of ribosomal DNA). Histology, however, had the advantage of assessing the impact of the parasite on the host. It was concluded that these parasites have the potential for significant ecological impacts, because of their high prevalence of infection and their ability to cause disease in some frogs.

Emerging myxosporean parasites of Australian frogs take a ride with fresh fruit transport.

BACKGROUND: The spread of wildlife pathogens into new geographical ranges or populations is a conservation concern for endangered species. Cystodiscus australis and Cystodiscus axonis are two species of myxosporean parasites infecting Australian frogs and tadpoles that have been recently recognised as important disease agents impacting amphibian conservation. Yet despite their importance to wildlife health, the mechanism of emergence for these parasites is unknown. We hypothesise that these parasites are capable of being accidentally translocated with their amphibian hosts in fresh produce (agricultural, horticultural and industrial) shipments into naïve environments and host populations. METHODS: We surveyed 33 Australian "Banana box" frogs from Sydney fruit markets during 2011 using faecal smears and multiplex species specific PCR on DNA isolated from frog faeces or using histopathology to demonstrate the presence of both C. australis and C. axonis. RESULTS: One of the "Banana box" frogs, the Dainty green tree frog (Litoria gracilenta) was positive for C. australis and C. axonis in its faeces and continuously shed the parasites for eight months. CONCLUSIONS: We present a possible mechanism for the emergence of Cystodiscus parasites and a non-invasive screening method to be used as a diagnostic test. In the future, vigilance and communication between wildlife managers/researchers and veterinarians will provide valuable information about these parasites, their host range and true distribution. This will aid risk management assessments for threatened populations within the range of Cystodiscus parasites and ultimately enhance conservation efforts.