An integrative taxonomy study reveals a rare new species of the genus Creptotrema (Trematoda: Allocreadiidae) in an endangered frog in South America.

During an ecological study with a near-endangered anuran in Brazil, the Schmidt's Spinythumb frog, Crossodactylus schmidti Gallardo, 1961, we were given a chance to analyze the gastrointestinal tract of a few individuals for parasites. In this paper, we describe a new species of an allocreadiid trematode of the genus Creptotrema Travassos, Artigas & Pereira, 1928, which possesses a unique trait among allocreadiids (i.e., a bivalve shell-like muscular structure at the opening of the ventral sucker); the new species represents the fourth species of allocreadiid trematode parasitizing amphibians. Besides, the new species is distinguished from other congeners by the combination of characters such as the body size, ventral sucker size, cirrus-sac size, and by having small eggs. DNA sequences through the 28S rDNA and COI mtDNA further corroborated the distinction of the new species. Phylogenetic analyses placed the newly generated sequences in a monophyletic clade together with all other sequenced species of Creptotrema. Genetic divergences between the new species and other Creptotrema spp. varied from 2.0 to 4.2% for 28S rDNA, and 15.1 to 16.8% for COI mtDNA, providing robust validation for the recognition of the new species. Even though allocreadiids are mainly parasites of freshwater fishes, our results confirm anurans as hosts of trematodes of this family. Additionally, we propose the reallocation of Auriculostoma ocloya Liquin, Gilardoni, Cremonte, Saravia, Cristóbal & Davies, 2022 to the genus Creptotrema. This study increases the known diversity of allocreadiids and contributes to our understanding of their evolutionary relationships, host-parasite relationships, and biogeographic history.

Phylogenetic position of Gorgoderina parvicava Travassos, 1922 (Digenea: Gorgoderidae), a parasite of Leptodactylus labyrinthicus (Spix, 1824) (Anura: Leptodactylidae) in Brazil.

During a parasite survey in Brazilian amphibians from São Paulo state, Brazil, Gorgoderina parvicava Travassos, 1922 was found in the urinary bladder (11 adult worms) and (five juvenile worms) in the kidneys of the pepper-frog Leptodactylus labyrinthicus (Spix, 1824). Parasites were examined by microscopy and 28S rDNA and COI gene were sequenced and analyzed for the molecular study. The phylogenetic reconstructions resulted in identical topologies with highly supported values in the nodes in most clades using Maximum likelihood and Bayesian inference methods and positioned G. parvicava in the subclade formed by species of subfamily Gorgoderinae parasitizing the urinary bladder of amphibians. Molecular phylogenetic data showed that this species is related to other species of Gorgoderina. In addition, new molecular data and the analyses of genetic distances provide extra comparative data, which can be applied in further investigations on the taxonomic status and the diversity among Gorgoderina spp. and host-parasite relationships.

First molecular assessment on Cosmocerca spp. from Brazilian anurans and description of a new species of Cosmocerca (Ascaridomorpha: Cosmocercoidea) from the white-spotted humming frog Chiasmocleis albopunctata (Boettger, 1885) (Anura: Microhylidae).

Cosmocerca spp. are common nematode parasites of amphibians. We provide herein molecular data for two species of Cosmocerca and describe a new species, Cosmocerca albopunctata n. sp., using light microscopy and molecular data (cytochrome c oxidase subunit 1 - COI mtDNA). Cosmocerca albopunctata n. sp. can be easily distinguished from other congeneric species by the combination of characteristics such as body size, length of spicules and gubernaculum, and the arrangements and number of caudal papillae (7 + 1:1 + 1:6). The phylogenetic results based on the partial COI mtDNA sequences clustered the new species in a monophyletic clade along with the other sequences of Cosmocerca spp. Therefore, our results contribute to the knowledge about the species diversity and genetic data for Cosmocerca spp. in the Neotropical region.

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.

Role of alpha-helix seven of Bacillus thuringiensis Cry1Ab delta-endotoxin in membrane insertion, structural stability, and ion channel activity.

Domain I of the Cry1Ab insecticidal toxic protein has seven alpha-helices and is considered to be involved in the ion channel activity. While other alpha-helices, particularly alpha-4 and alpha-5, have been extensively explored, the remaining alpha-helices have been slightly studied. Site-directed mutagenesis was used to generate mutations throughout sequences encoding the alpha-helix 7 to test its role in ion channel function. Every amino acid residue in alpha-helix 7 was mutated to alanine. Most resultant proteins, e.g., D225A, W226A, Y229A, N230A, R233A, R234A, D242A, and F247A yielded no protoxin or were sensitive to degradation by trypsin or Manduca sexta midgut juice. Other mutant proteins, R224A, R228A, and E235A, were resistant to degradation to the above proteases but were 8, 30, and 12 times less toxic to M. sexta, respectively, than the wild-type Cry1Ab. Circular dichroism spectroscopy indicated a very small change in the R228A spectrum, while R224A and E235A display the same spectrum as the wild-type protein. These three mutant proteins showed little differences from Cry1Ab when analyzed by saturation binding and competition binding kinetics with (125)I-labeled toxin or by surface plasmon resonance to M. sexta brush border membrane vesicles. More conservative amino acid substitutions were introduced into alpha-helix 7 residues: R228K, F232Y, E235Q, and F247Y. In comparison with wild-type Cry1Ab, mutant proteins R228K, F232Y, E235A, and E235Q selectively discriminate between K+ and Rb+, while R224A and R228A had reduced inhibition of short-circuit current for both ions, when analyzed by voltage clamping of M. sexta midguts.