Protozoan parasites of birds from the Tremembé formation (Oligocene of the Taubaté Basin), São Paulo, Brazil.

OBJECTIVE: To analyze the presence of protozoan parasites in bird coprolites from the Tremembé Formation (Oligocene of the Taubaté Basin). MATERIALS: Twenty avian coprolites embedded in pyrobituminous shale matrices. METHODS: Samples were rehydrated and subjected to spontaneous sedimentation. RESULTS: Paleoparasitological analyses revealed oocysts compatible with the Eimeriidae family (Apicomplexa) and one single Archamoebae (Amoebozoa) cyst. CONCLUSIONS: The present work increases the amount of information about the spread of infections throughout the Cenozoic Era and reveals that the Brazilian paleoavifauna played an important role in the Apicomplexa and Amoebozoa life cycles. SIGNIFICANCE: This is the first record of protozoans in avian coprolites from the Oligocene of Brazil. These findings can help in the interpretation of phylogenies of coccidian parasites of modern birds, as certain taxonomic characters observed in the Oligocene Protozoa characterize monophyletic groups in current molecular phylogenetic analyses. LIMITATIONS: None of the oocysts were sporulated; therefore, it is not possible to identify the morphotypes to genus or species. SUGGESTIONS FOR FURTHER RESEARCH: Our results create new perspectives related to biogeographic studies of the parasitic groups described and may improve the understanding of the temporal amplitude of parasitic evolutionary relationships between Protozoans and birds.

Oxysomatium brevispiculum n. sp. (Nematoda: Cosmocercidae) from Amphisbaena alba Linnaeus (Squamata, Amphisbaenidae) in the Cerrado Biome from Brazil.

In the present study, we describe Oxysomatium brevispiculum n. sp. (Ascaridida: Cosmocercidae) parasitizing Amphisbaena alba Linnaeus (Squamata: Amphisbaenidae) in the municipality of Uberlândia, Cerrado Biome, Minas Gerais, Brazil. Oxysomatium brevispiculum n. sp. differs from its congeners by having shorter spicules and by the number and arrangement of caudal papillae. The males of the new species have a precloacal unpaired papilla and can be easily distinguished from O. caucasicum in which this morphological trait is absent. Oxysomatium brevispiculum n. sp. differs from the other three species of the genus by the number and arrangement of caudal papillae, with 13 pairs + 1 unpaired precloacal papilla, arrangement 8+1:2:3, while O. brevicaudatum has 14-16 pairs +1 unpaired papilla, and 7-9+1:1:6 arrangement, O. petrolinensis has 16 pairs +1 unpaired papilla, and 8+1:1:7 arrangement, and O. dollfusi with 15-16 pairs +1 unpaired papilla, and 8-9+1:1:6 arrangement. Oxysomatium brevispiculum n. sp. is the fifth species of this genus, the second species in a Neotropical host, and the first species of this genus described in amphisbaenid hosts. In addition, the present study provides an identification key for the species of this genus.

Shaking the Tree: Multi-locus Sequence Typing Usurps Current Onchocercid (Filarial Nematode) Phylogeny.

During the past twenty years, a number of molecular analyses have been performed to determine the evolutionary relationships of Onchocercidae, a family of filarial nematodes encompassing several species of medical or veterinary importance. However, opportunities for broad taxonomic sampling have been scarce, and analyses were based mainly on 12S rDNA and coxI gene sequences. While being suitable for species differentiation, these mitochondrial genes cannot be used to infer phylogenetic hypotheses at higher taxonomic levels. In the present study, 48 species, representing seven of eight subfamilies within the Onchocercidae, were sampled and sequences of seven gene loci (nuclear and mitochondrial) analysed, resulting in the hitherto largest molecular phylogenetic investigation into this family. Although our data support the current hypothesis that the Oswaldofilariinae, Waltonellinae and Icosiellinae subfamilies separated early from the remaining onchocercids, Setariinae was recovered as a well separated clade. Dirofilaria, Loxodontofilaria and Onchocerca constituted a strongly supported clade despite belonging to different subfamilies (Onchocercinae and Dirofilariinae). Finally, the separation between Splendidofilariinae, Dirofilariinae and Onchocercinae will have to be reconsidered.

Physaloptera bainae n. sp. (Nematoda: Physalopteridae) parasitic in Salvator merianae (Squamata: Teiidae), with a key to Physaloptera species parasitizing reptiles from Brazil.

Physaloptera bainae n. sp. (Nematoda: Physalopteridae) from stomach of the large "tegú" lizard Salvator merianae (Squamata: Teiidae), collected in an ecological park that is part of the Atlantic Rainforest biome in Juiz de Fora, Minas Gerais, Brazil, is described. The new species which has females with didelphic uterus, is the only species of the genus exhibiting external teeth in the form of 4 spines that are organized in a cross-shaped pattern at the anterior apical end, with 2 minute adcloacal papillae on the anterior part of cloacal aperture in males and a large cuticular expansion at the anterior end of females. Moreover, P. bainae n. sp. can be differentiated from the other congeners by the combination of other features, e.g., number (23) and pattern of caudal papillae, spicule size (left 554-600; right 589-617) and vulval position (on the first third of body), and because the specimens are particularly large and robust. Physaloptera monodens is considered a junior synonym of P. obtusissima. In addition, a key to Physaloptera species parasitizing reptiles from Brazil is provided.

A new metastrongyloidean species (Nematoda) parasitizing pulmonary arteries of Puma (Herpailurus) yagouaroundi (É. Geoffroy, 1803) (Carnivora: Felidae) from Brazil.

Angiostrongylus felineus n. sp. (Nematoda, Metastrongyloidea), parasitic in Puma (Herpailurus) yagouaroundi (É. Geoffroy, 1803) (Carnivora, Felidae) from the municipality of Juiz de Fora, Minas Gerais state, Brazil, is described and illustrated herein. Angiostrongylus felineus n. sp. differs from all congeneric species by having the anterior extremity with accentuated cuticular expansion and by smaller size of spicules. This study describes for the first time a species of Angiostrongylus in a wild Felidae in Brazil.

Crenosoma brasiliense sp. n. (Nematoda: Metastrongyloidea) parasitic in lesser grison, Galictis cuja (Molina, 1782) (Carnivora, Mustelidae) from Brazil, with a key to species of Crenosoma Molin, 1861.

This study describes Crenosoma brasiliense (Nematoda, Metastrongyloidea), a new species parasitic in bronchi and bronchioles of Galictis cuja (Molina) (Carnivora, Mustelidae) from Brazil. This species differs from other 11 species of Crenosoma by having a cuticular projection at the distal end of the spicules, forming a prominent blade at the tip of the spicule, a vulval cuticular appendage with a triangular shape and prominent vulval lips. There are no previous records of species of Metastrongyloidea in G. cuja or species of Crenosoma in South America. Therefore, the new species represents the first host record and first geographical record of species of Crenosoma in South America.

A new type F Wolbachia from Splendidofilariinae (Onchocercidae) supports the recent emergence of this supergroup.

Wolbachia are vertically transmitted endosymbiotic bacteria of arthropods and onchocercid nematodes. It is commonly accepted that they co-evolved with their filarial hosts, and have secondarily been lost in some species. However, most of the data on the Wolbachia/Onchocercidae relationship have been derived from studies on two subfamilies, the Dirofilariinae and the Onchocercinae, which harbour parasites of humans and domestic animals. Within the last few years, analyses of more diverse material have suggested that some groups of Onchocercidae do not have Wolbachia, such as recently studied Splendidofilariinae from birds. This study takes advantage of the analysis of additional Splendidofilariinae, Rumenfilaria andersoni from a Finnish reindeer and Madathamugadia hiepei from a South African gecko, using PCR, immunohistochemical staining and whole-mount fluorescent analysis to detect Wolbachia and describe its strains. A DNA barcoding approach and phylogenetic analyses were used to investigate the symbiosis between Wolbachia and the Onchocercidae. A new supergroup F Wolbachia was demonstrated in M. hiepei, representing the first filarial nematode harbouring Wolbachia described in a non-mammalian host. In the adult, Wolbachia infects the female germline but not the hypodermis, and intestinal cells are also infected. The phylogenetic analyses confirmed a recent emergence of supergroup F. They also suggested several events of horizontal transmission between nematodes and arthropods in this supergroup, and the existence of different metabolic interactions between the filarial nematodes and their symbionts.

A new Physaloptera (Nematoda: Physalopteridae) parasite of Tupinambis merianae (Squamata: Teiidae) from southeastern Brazil.

Physaloptera tupinambae n. sp. (Nematoda: Physalopteridae) collected from the stomach of 1 Tupinambis merianae (Squamata: Teiidae) male lizard is described. Physaloptera tupinambae, which belongs to the didelphys group, is the only species in the genus that exhibits a bipartite internal tooth and in which the excretory pore is anterior to the deirids. The new species can be differentiated from the other congeners mainly based on the number and pattern of caudal papillae and the length and shape of spicules. Physaloptera murisbrasiliensis and Physaloptera clausa orientalis are the only physalopterids with the same number of caudal papillae as for P. tupinambae, but they differ from the new species mainly in length and shape of spicules. Based on morphological analysis, the speciation process among Physaloptera probably occurred via host capture, and P. tupinambae occupies an intermediate phylogenetic position in the genus. Most likely, the new parasite was acquired by the lizard through ingestion of an intermediate host. Finally, Physaloptera calotisi, Physaloptera funambuli, Physaloptera guptae, Physaloptera indica, Physaloptera johnsoni, Physaloptera kherai, and Physaloptera thaparus are moved to Abbreviata.

New insights into the evolution of Wolbachia infections in filarial nematodes inferred from a large range of screened species.

BACKGROUND: Wolbachia are intriguing symbiotic endobacteria with a peculiar host range that includes arthropods and a single nematode family, the Onchocercidae encompassing agents of filariases. This raises the question of the origin of infection in filariae. Wolbachia infect the female germline and the hypodermis. Some evidences lead to the theory that Wolbachia act as mutualist and coevolved with filariae from one infection event: their removal sterilizes female filariae; all the specimens of a positive species are infected; Wolbachia are vertically inherited; a few species lost the symbiont. However, most data on Wolbachia and filaria relationships derive from studies on few species of Onchocercinae and Dirofilariinae, from mammals. METHODOLOGY/PRINCIPAL FINDINGS: We investigated the Wolbachia distribution testing 35 filarial species, including 28 species and 7 genera and/or subgenera newly screened, using PCR, immunohistochemical staining, whole mount fluorescent analysis, and cocladogenesis analysis. (i) Among the newly screened Onchocercinae from mammals eight species harbour Wolbachia but for some of them, bacteria are absent in the hypodermis, or in variable density. (ii) Wolbachia are not detected in the pathological model Monanema martini and in 8, upon 9, species of Cercopithifilaria. (iii) Supergroup F Wolbachia is identified in two newly screened Mansonella species and in Cercopithifilaria japonica. (iv) Type F Wolbachia infect the intestinal cells and somatic female genital tract. (v) Among Oswaldofilariinae, Waltonellinae and Splendidofilariinae, from saurian, anuran and bird respectively, Wolbachia are not detected. CONCLUSIONS/SIGNIFICANCE: The absence of Wolbachia in 63% of onchocercids, notably in the ancestral Oswaldofilariinae estimated 140 mya old, the diverse tissues or specimens distribution, and a recent lateral transfer in supergroup F Wolbachia, modify the current view on the role and evolution of the endosymbiont and their hosts. Further genomic analyses on some of the newly sampled species are welcomed to decipher the open questions.